Formulations for hydrophobic pharmaceutical agents

ABSTRACT

The present invention features formulations, including liquid, semi-solid or solid pharmaceutical formulations, that improve the oral bioavailability of hydrophobic pharmaceutical agents, such as quinazoline-, nitrothiazole-, and indolinone-based compounds. Also featured are formulations for parenteral delivery of such hydrophobic pharmaceutical agents, as well as methods of making and using both types of formulations.

STATEMENT OF RELATED APPLICATIONS

This application is related to U.S. patent application Ser. No.60/041,251, filed Mar. 18, 1997 and U.S. patent application Ser. No.60/039,870, filed Mar. 5, 1997, both of which are incorporated herein byreference in their entirety, including any drawings.

FIELD OF THE INVENTION

The present invention provides liquid, semi-solid or solidpharmaceutical formulations that improve the oral bioavailability ofhydrophobic pharmaceutical agents, such as indolinone-, quinazoline-,and nitrothiazole-based compounds. Also provided are formulations forparenteral delivery of such hydrophobic pharmaceutical agents, as wellas methods of making and using both types of formulations.

BACKGROUND OF THE INVENTION

The following description of the background of the invention is providedto aid in understanding the invention, but is not admitted to describeor constitute prior art to the invention.

Various methods are available for administering therapeutic agents to apatient. Such methods include, parenteral, oral, ocular, nasal, topical,and transmucosal administration. Variations of these different types ofadministrations exist. For example, parenteral administration includesintravenous, subcutaneous, intraperitoneal, intramuscular, andintramedullary injection. The chosen mode of administration should takeinto account the nature of the therapeutic compound and the illnessbeing treated.

Certain potential pharmaceuticals are hydrophobic and typically havevery low aqueous solubility and hence low oral bioavailability.Different techniques concerned with solubilizing hydrophobic compoundsinclude those described by Praveen et al., U.S. Pat. No. 5,314,685, andFernandes et al., U.S. Pat. No. 4,992,271, both of which areincorporated by reference herein in their entirety including any figuresand drawings.

One measure of the potential usefulness of an oral formulation of a newpharmaceutical agent is the bioavailability observed after oraladministration of the formulation. Various factors can affect the oralbioavailability of the drug. These factors include aqueous solubility,drug absorption throughout the gastrointestinal tract, dosage strength,and first pass effect. Aqueous solubility is one of the most importantfactors. The oral bioavailability of an aqueous solution formulation ofa drug is generally used as the standard or the ideal bioavailabilityagainst which other oral formulations are measured. Formulations ofdrugs that increase the relative bioavailability of the drug as comparedto an aqueous solution are desirable, especially with hydrophobiccompounds.

SUMMARY OF THE INVENTION

The present invention features formulations (including formulations fororal administration as well as parenteral administration) forhydrophobic pharmaceutical agents, such as indolinone-, quinazoline-, ornitrothiazole-based compounds. Such formulations have advantageoussolubility characteristics which allow for administration of hydrophobicpharmaceutical agents, such as indolinone-, quinazoline-, ornitrothiazole-based compounds, for pharmaceutical testing and therapy.Not only do such formulations overcome the solubility problems that havepreviously plagued the art, they have also been shown to produce atherapeutic effect in test animals.

Thus, a first aspect of the present invention features a formulationcomprising: (a) one or more hydrophobic pharmaceutical agents, where theagents are independently selected from the group consisting ofquinazoline-, nitrothiazole-, and indolinone-based compounds; (b) one ormore polyoxyhydrocarbyl compounds; and (c) one or more pharmaceuticallyacceptable surfactants.

It is anticipated that the one or more hydrophobic pharmaceutical agentsmay include a combination of nitrothiazole-based compounds withquinazoline-based compounds, or nitrothiazole-based compounds withindolinone-based compounds, or quinazoline-based compounds withindolinone-based compounds. In addition, the one or more hydrophobicpharmaceutical agents may include a combination of indolinone-basedcompounds, for example3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-amino)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-chloro)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, and3-[(3-methylthiophenyl-5-yl)methylene]-(4-methyl)-2-indolinone. Anotherpossiblity is that the one or more hydrophobic pharmaceutical agents mayinclude a combination of quinazoline-based compounds, for example4-(3-bromophenyl)-6,7-dimethoxyquinazoline and4-(3-chlorophenyl)-6,7-dimethoxyquinazoline. Or alternatively, the oneor more hydrophobic pharmaceutical agents may include a combination ofnitrothiazole-based compounds, for example some combination of2-methyl-5-[(5-nitrothiazol-2-yl)mercapto]-1,3,4-thiadiazole;1-benzyl-5-[(5-nitrothiazol-2-yl)mercapto]tetrazole;2-[(5-nitrothiazol-2-yl)mercapto]-5-t-butyl-1,2,4-triazole;3-[(5-nitrothiazol-2-yl)mercapto]-5-(thien-2-yl)-1,2,4-triazole;3-[(5-nitrothiazol-2-yl)mercapto]-5-phenyl-1,2,4-triazole; and4-allyl-3-hydroxy-5-[(5-nitrothiazole-2-yl)mercapto]-1,2,4-triazole.

The term “hydrophobic pharmaceutical agent” as used herein refers tocompounds having a greater solubility in organic solvents of lowpolarity, such as long chain alcohols, than in aqueous solution.“Hydrophobic” means “water-hating” and is used herein to indicate weaklysoluble in water and soluble in non-polar solvents. The formulationsdescribed by the present invention facilitate solubilization ofhydrophobic compounds which readily dissolve in alcohols. Preferably,the hydrophobic compound is insoluble in aqueous solution. Morepreferably, the compound has similar solubility characteristics inalcohols and aqueous solution to quinazoline-, nitrothiazole-, andindolinone-based compounds.

The term “compound” refers to the compound or a pharmaceuticallyacceptable salt, ester, amide, prodrug, isomer, or metabolite, thereof.

The term “pharmaceutically acceptable salt” refers to a formulation of acompound that does not abrogate the biological activity and propertiesof the compound. Pharmaceutical salts can be obtained by reacting acompound of the invention with inorganic or organic acids such ashydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicylic acid and the like, or with inorganicor organic bases such as sodium hydroxide, potassium hydroxide, ammoniumhydroxide, calcium hydroxide, choline, n-methyl glucamine, diethylamine,procaine and the like.

The term “prodrug” refers to an agent that is converted into the parentdrug in vivo. Prodrugs may be easier to administer than the parent drugin some situations. For example, the prodrug may be bioavailable by oraladministration but the parent is not, or the prodrug may improvesolubility to allow for intravenous administration.

The term “polarity” as used herein refers to the dipole of a molecule. A“dipole” is two equal and opposite charges separated in space. Amolecule is polar if it has a dipole, i.e. if the center of negativecharge does not coincide with the center of positive charge. The dipolemoment of a molecule, is equal to the magnitude of the charge multipliedby the distance between the centers of charge. It is possible to measurethe dipole moments of molecules by methods well-known in the art. A lowdipole moment indicates low polarity.

The term “quinazoline-based compound” refers to a quinazoline organiccompound substituted with chemical substituents. Quinazoline compoundsare of the general structure:

The term “substituted” refers to compounds of the invention that arederivatized with any number of chemical substituents, typicallyreplacing one or more of the hydrogen atoms present in the compound'sgeneral structure.

The term “nitrothiazole-based compound” refers to a nitrothiazoleorganic compound substituted with chemical substituents. Nitrothiazolecompounds are of the general structure:

The term “indolinone-based compound” refers to a indolinone organiccompound substituted with chemical substituents. Indolinone compoundsare of the general structure:

The term “polyoxyhydrocarbyl compound” as used herein refers to a watersoluble carbohydrate such as glucose, sucrose, maltotriose, and thelike; water soluble carbohydrate derivatives such as gluconic acid andmannitol, and oligosaccharides; and water soluble polymers such aspolyvinylpyrrolidone, poly(vinyl alcohol), and in particular, polyetherssuch as other polyoxyalkylenes including poly(ethylene glycol), or otherwater soluble mixed oxyalkylene polymers, and the polymeric form ofethylene glycol. Although polyoxyhydrocarbyl compounds preferablycontain more than one carbon, oxygen, and hydrogen atom, some moleculessuch as poly(ethyleneimine) are also included.

A particularly preferred class of solubilizing polyoxyhydrocarbylmoieties comprises poly(ethylene glycol) (PEG) and PEG derivatives, suchas PEG monomethyl ether. Other suitable PEG derivatives includePEG-silicon derived ethers. Many of these polymers are commerciallyavailable in a variety of molecular weights. Others may be convenientlyprepared from commercially available materials, such as by coupling ofamino-PEG moiety to a haloalkyl silyl or silane moiety.

Suitable PEGs may vary in molecular weight from about 200 g/mol to about20,000 g/mol or more, more preferably 200 g/mol to 5,000 g/mol, evenmore preferably 250 g/mol to 1,000 g/mol, and most preferably 250 g/molto 500 g/mol. The choice of a particular molecular weight may depend onthe particular hydrophobic pharmaceutical agent chosen and its molecularweight and degree of hydrophobicity, as well as the particularapplication for which the formulation is to be used.

The term “pharmaceutically acceptable surfactant” as used herein refersto a compound that can solubilize hydrophobic compounds into aqueoussolutions. Preferably for parenteral formulations, the surfactant is anon-ionic surfactant. Examples of pharmaceutically acceptablesurfactants include POLYSORBATE 80® and other polyoxyethylene sorbitanfatty acid esters, glyceryl monooleate, polyvinyl alcohol, ethyleneoxide copolymers such as PLURONIC™ (a polyether) and TETRONIC™ (BASF),polyol moieties, and sorbitan esters. Preferably ethoxylated castoroils, such as CREMOPHOR EL®, are used for the formulation of hydrophobicpharmaceutical agents, such as indolinone-, quinazoline-, andnitrothiazole-based compounds.

The term “ethoxylated castor oil” as used herein refers to castor oilthat is modified with at least one oxygen containing moiety. Inparticular the term refers to castor oil comprising at least one ethoxylmoiety.

Further, the term “pharmaceutically acceptable surfactant” as usedherein in reference to oral formulations, includes pharmaceuticallyacceptable non-ionic surfactants (for examplepolyoxyethylenepolypropylene glycol, such as POLOXAMER® 68 (BASF Corp.)or a mono fatty acid ester of polyoxyethylene (20) sorbitan monooleate(TWEEN® 80), polyoxyethylene (20) sorbitan monostearate (TWEEN® 60),polyoxyethylene (20) sorbitan monopalmitate (TWEEN® 40), polyoxyethylene(20) sorbitan monolaurate (TWEEN® 20) and the like); polyoxyethylenecastor oil derivatives (for example,polyoxyethyleneglyceroltriricinoleate or polyoxyl 35 castor oil(CREMOPHOR® EL, BASF Corp.), polyoxyethyleneglycerol oxystearate(CREMOPHOR® RH 40 (polyethyleneglycol 40 hydrogenated castor oil) orCREMOPHOR® RH 60 (polyethyleneglycol 60 hydrogenated castor oil), BASFCorp.) and the like); or a pharmaceutically acceptable anionicsurfactant.

The term “pharmaceutically acceptable” or “pharmaceutical” as usedherein refers to solutions or components of the formulation that do notprevent the therapeutic compound from exerting a therapeutic effect anddo not cause unacceptable adverse side effects. Examples ofpharmaceutically acceptable reagents are provided in The United StatesPharmacopeia The National Formulary, United States PharmacopeialConvention, Inc., Rockville, Md. 1990 and FDA Inactive Ingredient Guide1990, 1996 issued by the Division of Drug Information Resources (bothare hereby incorporated by reference herein, including any drawings).Unacceptable side effects vary for different diseases. Generally, themore severe the disease the more toxic effects which will be tolerated.Unacceptable side effects for different diseases are known in the art.

In preferred embodiments of the invention, the hydrophobicpharmaceutical agent is a quinazoline-based compound of formula I,

where R₁, R₂, R₃, R₄, R₅ and R₆ are independently selected from thegroup consisting of: (i) hydrogen; (ii) saturated or unsaturated alkyl;(iii) an aryl optionally substituted with one, two, or threesubstituents independently selected from the group consisting of alkyl,alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester moieties;(iv) an amine of formula —NX₂X₃, where X₂ and X₃ are independentlyselected from the group consisting of hydrogen, saturated or unsaturatedalkyl, and homocyclic or heterocyclic ring moieties; (v) halogen ortrihalomethyl; (vi) a ketone of formula —CO—X₄, where X₄ is selectedfrom the group consisting of alkyl and homocyclic or heterocyclic ringmoieties; (vii) a carboxylic acid of formula —(X₅)_(n)—COOH or ester offormula —(X₆)_(n)—COO—X₇, where X₅, X₆, and X₇ and are independentlyselected from the group consisting of alkyl and homocyclic orheterocyclic ring moieties and where n is 0 or 1; (viii) an alcohol offormula (X₈)_(n)—OH or an alkoxy moiety of formula —(X₈)_(n)—O—X₉, whereX₈ and X₉ are independently selected from the group consisting ofsaturated or unsaturated alkyl and homocyclic or heterocyclic ringmoieties, wherein said ring is optionally substituted with one or moresubstituents independently selected from the group consisting of alkyl,alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester and wheren is 0 or 1; (ix) an amide of formula —NHCOX₁₀, where X₁₀ is selectedfrom the group consisting of alkyl, hydroxyl, and homocyclic orheterocyclic ring moieties, wherein said ring is optionally substitutedwith one or more substituents independently selected from the groupconsisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, nitro,and ester; (x) —SO₂NX₁₁X₁₂, where X₁₁ and X₁₂ are selected from thegroup consisting of hydrogen, alkyl, and homocyclic or heterocyclic ringmoieties; (xi) a homocyclic or heterocyclic ring moiety optionallysubstituted with one, two, or three substituents independently selectedfrom the group consisting of alkyl, alkoxy, halogen, trihalomethyl,carboxylate, nitro, and ester moieties; (xii) an aldehyde of formula—CO—H; (xiii) a sulfone of formula —SO₂—X₁₃, where X₁₃ is selected fromthe group consisting of saturated or unsaturated alkyl and homocyclic orheterocyclic ring moieties; and (xiv) a nitro of formula —NO₂.

The term “saturated alkyl” refers to an alkyl moiety that does notcontain any alkene or alkyne moieties. The alkyl moiety may be branchedor non-branched.

The term “unsaturated alkyl” refers to an alkyl moiety that contains atleast one alkene or alkyne moiety. The alkyl moiety may be branched ornon-branched.

The term “aryl” refers to an aromatic group which has at least one ringhaving a conjugated pi electron system and includes both carbocyclicaryl (e.g. phenyl) and heterocyclic aryl groups (e.g. pyridine). Theterm “carbocyclic” refers to a compound which contains one or morecovalently closed ring structures, and where the atoms forming thebackbone of the ring are all carbon atoms. The term thus distinguishescarbocyclic from “heterocyclic” rings in which the ring backbonecontains at least one atom which is different from carbon. The term“heteroaryl” refers to an aryl group which contains at least oneheterocyclic ring.

The term “amine” refers to a chemical moiety of formula NR₁R₂ where R₁and R₂ are independently selected from the group consisting of hydrogen,saturated or unsaturated alkyl, and homocyclic or heterocyclic ringmoieties, where the ring is optionally substituted with one or moresubstituents independently selected from the group consisting of alkyl,halogen, trihalomethyl, carboxylate, nitro, and ester moieties.

The term “halogen” refers to an atom selected from the group consistingof fluorine, chlorine, bromine, and iodine.

The term “ketone” refers to a chemical moiety with formula—(R)_(n)—CO—R′, where R and R′ are selected from the group consisting ofsaturated or unsaturated alkyl and homocyclic or heterocyclic ringmoieties and where n is 0 or 1.

The term “carboxylic acid” refers to a chemical moiety with formula—(R)_(n)—COOH, where R is selected from the group consisting ofsaturated or unsaturated alkyl and homocyclic or heterocyclic ringmoieties, and where n is 0 or 1.

The term “alcohol” refers to a chemical substituent of formula —ROH,where R is selected from the group consisting of saturated orunsaturated alkyl, and homocyclic or heterocyclic ring moieties, wherethe ring moiety is optionally substituted with one or more substituentsindependently selected from the group consisting of alkyl, halogen,trihalomethyl, carboxylate, nitro, and ester moieties.

The term “ester” refers to a chemical moiety with formula—(R)_(n)—COOR′, where R and R′ are independently selected from the groupconsisting of saturated or unsaturated alkyl and homocyclic orheterocyclic ring moieties and where n is 0 or 1.

The term “alkoxy” refers to a chemical substituent of formula —OR, whereR is hydrogen or a saturated or unsaturated alkyl moiety.

The term “amide” refers to a chemical substituent of formula —NHCOR,where R is selected from the group consisting of hydrogen, alkyl,hydroxyl, and homocyclic or heterocyclic ring moieties, where the ringis optionally substituted with one or more substituents independentlyselected from the group consisting of alkyl, halogen, trihalomethyl,carboxylate, nitro, or ester.

The term “aldehyde” refers to a chemical moiety with formula—(R)_(n)—CHO, where R is selected from the group consisting of saturatedor unsaturated alkyl and homocyclic or heterocyclic ring moieties andwhere n is 0 or 1.

The term “sulfone” refers to a chemical moiety with formula —SO₂—R,where R is selected from the group consisting of saturated orunsaturated alkyl and homocyclic or heterocyclic ring moieties.

In preferred embodiments, the hydrophobic pharmaceutical agent is aquinazoline-based compound of formula II:

where R₁, R₂, and R₃ are selected from the group consisting of halogen,trihalomethyl, cyano, methoxy, and hydrogen. Most preferably, thequinazoline-based compound is4-(3-bromophenyl)-6,7-dimethoxyquinazoline.

In other preferred embodiments of the invention, the hydrophobicpharmaceutical agent is a nitrothiazole-based compound of formula III,

where A represents (i) a substituted or unsubstituted monocyclic five-or six-membered ring having 1 to 4 hetero ring atoms, at least one ofwhich is nitrogen, the remainder of which are selected from the groupconsisting of nitrogen, oxygen and sulfur, where examples of such ringsinclude, but are not limited to, pyridine, pyrrole, imidazole, thiazole,isothiazole, isoxazole, furazan, pyrrolidine, piperidine, imidazolidine,piperazine, oxazole, tetrazole, pyrazole, triazole, oxadiazole,thiodiazole; (ii) a substituted or unsubstituted monocyclic or fusedbicyclic six- to ten-membered ring having 1 to 4 hetero ring atoms, oneof which is nitrogen and the remainder of which are selected from thegroup consisting of nitrogen, oxygen and sulfur, where such ringsinclude, .but are not limited to, indole, quinoxaline, quinazoline,quinoline, isoquinoline, purine; or (iii) a substituted or unsubstitutedmonocyclic or fused polycyclic saturated or unsaturated ring havingthree to 15 atoms, which are selected from the group consisting ofcarbon, sulfur, nitrogen and oxygen.

The heterocyclic rings defined above may be saturated or unsaturated.The unsaturated rings or heteroaromatic group may, if desired, bear oneor more substituents which do not substantially adversely affect theactivity of the compound of formula II. Exemplary of such substituentsare alkyl, alkoxy, phenoxy, alkenyl, alkynyl, phenylalkyl, hydroxyalkyl,haloalkyl, aryl, arylalkyl, alkyloxy, alkylthio, alkenylthio,phenylalkylthio, hydroxyalkyl-thio, alkylthiocarbamylthio, phenyl,cyclohexyl, pyridyl, piperidinyl, alkylamino, amino, nitro, mercapto,cyano, hydroxyl, a halogen atom, an oxygen atom (forming a ketone orN-oxide) or a sulphur atom (forming a thione).

The terms “alkenyl” and “alkynylu as used herein refer to straight orbranched chain hydrocarbon groups having from 2 to 10 carbons andunsaturated by a double or triple bond, respectively, such as vinyl,allyl, propargyl, 1-methylvinyl, but-1-enyl, but-2-enyl, but-2-ynyl, 1methylbut-2-enyl, pent-1-enyl, pent-3-enyl, 3-methylbut-1-ynyl,1,1-dimethylallyl, hex-2-enyl and 1-methyl-1-ethylallyl.

The term “phenylalkyl” refers to the aforementioned alkyl groupssubstituted by a phenyl group. Examples of phenylalkyl groups include,but are not limited to, benzyl, phenethyl, phenopropyl, 1-benzylethyl,phenobutyl and 2-benzylpropyl. The term “hydroxy-alkyl” refers to theaforementioned alkyl groups substituted by a single hydroxyl group.Examples of hydroxyalkyl goups include, but are not limited to,2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 4-hydroxybutyl,1-hydroxybutyl and 6-hydroxyhexyl.

The terms “alkylthio, alkenylthio, alkynylthio, alkylthio,hydroxy-alkylthio and phenyl-alkylthio” as used herein refer to theaforementioned alkyl, alkenyl, alkynyl, hydroxy-alkyl and phenyl-alkylgroups linked through a sulfur atom to the compounds of the presentinvention.

In yet other preferred embodiments, the hydrophobic pharmaceutical agentis a nitrothiazole-based compound of formula IV:

where R₁, R₂, and R₃ are independently selected from the groupconsisting of: (i) hydrogen; (ii) saturated or unsaturated alkyl; (iii)an aryl optionally substituted with one, two, or three substituentsindependently selected from the group consisting of alkyl, alkoxy,halogen, trihalomethyl, carboxylate, nitro, and ester moieties; (iv) anamine of formula —NX₂X₃, where X₂ and X₃ are independently selected fromthe group consisting of hydrogen, saturated or unsaturated alkyl, andhomocyclic or heterocyclic ring moieties; (v) halogen or trihalomethyl;(vi) a ketone of formula —CO—X₄, where X₄ is selected from the groupconsisting of alkyl and homocyclic or heterocyclic ring moieties; (vii)a carboxylic acid of formula —(X₅)_(n)—COOH or ester of formula—(X₆)_(n)—COO—X₇, where X₅, X₆, and X₇ and are independently selectedfrom the group consisting of alkyl and homocyclic or heterocyclic ringmoieties and where n is 0 or 1; (viii) an alcohol of formula (X₈)_(n)—OHor an alkoxy moiety of formula —(X₈)_(n)—O—X₉, where X₈ and X₉ areindependently selected from the group consisting of saturated orunsaturated alkyl and homocyclic or heterocyclic ring moieties, whereinsaid ring is optionally substituted with one or more substituentsindependently selected from the group consisting of alkyl, alkoxy,halogen, trihalomethyl, carboxylate, nitro, and ester and where n is 0or 1; (ix) an amide of formula —NHCOX₁₀, where X₁₀ is selected from thegroup consisting of alkyl, hydroxyl, and homocyclic or heterocyclic ringmoieties, wherein said ring is optionally substituted with one or moresubstituents independently selected from the group consisting of alkyl,alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester; (x)—SO₂NX₁₁X₁₂, where X₁₁ and X₁₂ are selected from the group consisting ofhydrogen, alkyl, and homocyclic or heterocyclic ring moieties; (xi) ahomocyclic or heterocyclic ring moiety optionally substituted with one,two, or three substituents independently selected from the groupconsisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, nitro,and ester moieties; (xii) an aldehyde of formula —CO—H; (xiii) a sulfoneof formula —SO₂—X₁₃, where X₁₃ is selected from the group consisting ofsaturated or unsaturated alkyl and homocyclic or heterocyclic ringmoieties; and (xiv) a nitro of formula —NO₂.

In yet other preferred embodiments of the invention, the hydrophobicpharmaceutical agent is a nitrothiazole-based compound of formula V:

or a pharmaceutically acceptable salt thereof, wherein R₁ and R₂ areindependently selected from the group consisting of: (i) hydrogen; (ii)saturated or unsaturated alkyl; (iii) an aryl optionally substitutedwith one, two, or three substituents independently selected from thegroup consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate,nitro, and ester moieties; (iv) an amine of formula —NX₂X₃, where X₂ andX₃ are independently selected from the group consisting of hydrogen,saturated or unsaturated alkyl, and homocyclic or heterocyclic ringmoieties; (v) halogen or trihalomethyl; (vi) a ketone of formula —CO—X₄,where X₄ is selected from the group consisting of alkyl and homocyclicor heterocyclic ring moieties; (vii) a carboxylic acid of formula—(X₅)_(n)—COOH or ester of formula —(X₆)_(n)—COO—X₇, where X₅, X₆, andX₇ and are independently selected from the group consisting of alkyl andhomocyclic or heterocyclic ring moieties and where n is 0 or 1; (viii)an alcohol of formula (X₈)_(n)—OH or an alkoxy moiety of formula—(X₈)_(n)—O—X₉, where X₈ and X₉ are independently selected from thegroup consisting of saturated or unsaturated alkyl and homocyclic orheterocyclic ring moieties, wherein said ring is optionally substitutedwith one or more substituents independently selected from the groupconsisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, nitro,and ester and where n is 0 or 1; (ix) an amide of formula —NHCOX₁₀,where X₁₀ selected from the group consisting of alkyl, hydroxyl, andhomocyclic or heterocyclic ring moieties, wherein said ring isoptionally substituted with one or more substituents independentlyselected from the group consisting of alkyl, alkoxy, halogen,trihalomethyl, carboxylate, nitro, and ester; (x) —SO₂NX₁₁X₁₂, where X₁₁and X₁₂ are selected from the group consisting of hydrogen, alkyl, andhomocyclic or heterocyclic ring moieties; (xi) a homocyclic orheterocyclic ring moiety optionally substituted with one, two, or threesubstituents independently selected from the group consisting of alkyl,alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester moieties;(xii) an aldehyde of formula —CO—H; (xiii) a sulfone of formula—SO₂—X₁₃, where X₁₃ is selected from the group consisting of saturatedor unsaturated alkyl and homocyclic or heterocyclic ring moieties; and(xiv) a nitro of formula —NO₂.

In particularly preferred embodiments, the nitrothiazole-based compoundis selected from the group consisting of:2-methyl-5-[(5-nitrothiazol-2-yl)mercapto]-1,3,4-thiadiazole;1-benzyl-5-[(5-nitrothiazol-2-yl)mercapto]tetrazole;2-[(5-nitrothiazol-2-yl)mercapto]-5-t-butyl-1,2,4-triazole;3-[(5-nitrothiazol-2-yl)mercapto]-5-(thien-2-yl)-1,2,4-triazole;3-[(5-nitrothiazol-2-yl)mercapto]-5-phenyl-1,2,4-triazole; and4-allyl-3-hydroxy-5-[(5-nitrothiazole-2-yl)mercapto]-1,2,4-triazole.

In yet other preferred embodiments of the invention, the hydrophobicpharmaceutical agent is a indolinone-based compound of formula VI:

where R₁, R₂, R₃, and R₄ are selected from the group consisting ofhydrogen, trihalomethyl, hydroxyl, amine, thioether, cyano, alkoxy,alkyl, amino, bromo, fluoro, chloro, iodo, mercapto, thio, cyanoamido,alkylthio, aryl, heteroaryl, carboxyl, ester, oxo, alkoxycarbonyl,alkenyl, alkoxy, nitro, alkoxyl, and amido moieties; and R₅ is anoptionally substituted aryl or heteroaryl cyclic moiety.

Preferably, the indolinone-based compound has a structure of formula VI,where R₁, R₂, R₃, and R₄ are selected from the group consisting ofhydrogen, halogen, alkyl, and alkoxy; and where R₅ is a pyrrolyl orthiophenyl moiety optionally substituted with moieties selected from thegroup consisting of hydrogen, halogen, alkyl, and alkoxy.

More preferably, the indolinone-based compound is selected from thegroup consisting of3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-amino)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-chloro)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, and3-[(3-methylthiophenyl-5-yl)methylene]-(4-methyl)-2-indolinone. Mostpreferably, the indolinone-based compound is3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone.

In some embodiments of the invention, the one or more polyoxyhydrocarbylcompounds are independently selected from the group consisting of: watersoluble carbohydrates, water soluble carbohydrate derivatives,polypeptides, water soluble polymers, water soluble mixed oxyalkylenepolymers, and the polymeric form of ethylene glycol. Preferably, the oneor more polyoxyhydrocarbyl compounds are poly(ethylene glycol) (PEG) orPEG derivatives. More preferably, PEG may vary in molecular weight fromabout 200 daltons to about 20,000 daltons.

In other embodiments of the invention, the one or more surfactants areone or more non-ionic surfactants. Preferably, the one or moresurfactants are independently selected from the group consisting of:polyoxyethylene sorbitan fatty acid esters, glyceryl monooleate,polyvinyl alcohol, ethylene oxide copolymers, polyol moieties, andsorbitan esters. More preferably, the one or more surfactants are one ormore ethoxylated castor oils. Most preferably, the ethoxylated castoroil is CREMOPHOR EL®.

In preferred embodiments of the invention the formulation also includesone or more pharmaceutically acceptable alcohols. Preferably, the one ormore alcohols are independently selected from the group consisting ofethanol, benzyl alcohol, propylene glycol, 2-(2-ethoxyethoxy)ethanol,and glycerol. Most preferably, the alcohols are ethanol and benzylalcohol.

The term “pharmaceutically acceptable alcohol” as used herein refers toalcohols which are liquids at about room temperature (approximately 20°C.). These include propylene glycol, ethanol, 2-(2-ethoxyethoxy)ethanol(TRANSCUTOL®, Gattefosse, Westwood, N.J. 07675), benzyl alcohol, andglycerol.

The formulation should be dissolved in a sufficient amount of apharmaceutically acceptable aqueous solution prior to patientadministration to avoid toxic effects due to the alcohol content. Theadded amount of a pharmaceutically acceptable aqueous solution should besufficient to avoid hemolysis. Examples of suitable pharmaceuticallyacceptable aqueous solutions such as WFI (water for injection) andsolutions containing isotonic saline are known in the art.Pharmaceutically acceptable aqueous solutions include 0.45% N saline,WFI (water for injection), D5W (5% dextrose in water), and D5W 0.450% Nsaline.

In other embodiments of the invention, when the hydrophobicpharmaceutical agent is an indolinone-based compound substituted withone or more carboxyl moieties, the formulation further comprises water.

In preferred embodiments of the invention, the formulation comprises (a)0.1 to 100 mg/mL of an indolinone-based compound of formula VI, whereR₁, R₂, R₃, and R₄ are selected from the group consisting of hydrogen,trihalomethyl, hydroxyl, amine, thioether, cyano, alkoxy, alkyl, amino,bromo, fluoro, chloro, iodo, mercapto, thio, cyanoamido, alkylthio,aryl, heteroaryl, carboxyl, ester, oxo, alkoxycarbonyl, alkenyl, alkoxy,nitro, alkoxyl, and amido moieties; and R₅ is an optionally substitutedaryl or heteroaryl cyclic moiety; (b) 0.01 to 10 g/mL PEG-400; (c) 0.01to 1 g/mL ethanol; (d) 0.001 to 1 g/mL benzyl alcohol; and (e) 0.01 to10 g/mL ethoxylated castor oil.

The term “PEG-400” as used herein refers to a polymeric form of ethyleneglycol, polyethylene glycol (PEG), which has an average molecular weightof 400 grams/mole.

In other preferred embodiments the formulation comprises (a) about 5.0mg/mL of an indolinone-based compound of formula VI, where R₁, R₂, R₃,and R₄ are selected from the group consisting of hydrogen,trihalomethyl, hydroxyl, amine, thioether, cyano, alkoxy, alkyl, amino,bromo, fluoro, chloro, iodo, mercapto, thio, cyanoamido, alkylthio,aryl, heteroaryl, carboxyl, ester, oxo, alkoxycarbonyl, alkenyl, alkoxy,nitro, alkoxyl, and amido moieties; and R₅ is an optionally substitutedaryl or heteroaryl cyclic moiety; (b) about 0.35 g/mL PEG-400 (c) about0.114 g/mL ethanol; (d) about 0.02 g/mL benzyl alcohol; and (e) about0.25 g/mL CREMOPHOR EL®.

In other preferred embodiments, the invention relates to formulationscomprising an indolinone-based compound of formula VI, where R₁, R₂, R₃,and R₄ are selected from the group consisting of hydrogen, halogen,alkyl, and alkoxy; and where R₅ is a pyrrolyl or thiophenyl moietyoptionally substituted with moieties selected from the group consistingof hydrogen, halogen, alkyl, and alkoxy.

In still other preferred embodiments, the invention relates toformulations comprising an indolinone-based compound selected from thegroup consisting of3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-amino)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-chloro)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, and3-[(3-methylthiophenyl-5-yl)methylene]-(4-methyl)-2-indolinone.

A highly preferred embodiment of the invention relates to a formulationcomprising about 4.5 mg/mL of the indolinone-based compound,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, (b) about 45t w/vPEG-400, (c) about 31.5% w/v CREMOPHOR EL®, (d) about 2% w/v benzylalcohol, and (e) about 9.5% w/v ethanol.

The formulations are stated to comprise the elements described herein,meaning that the formulations can include other components. The solutioncontaining the hydrophobic pharmaceutical agents is preferably adjustedto a pH where the compounds are stable. The pH is preferably adjusted tobetween 2 and 7. The pH can be adjusted using pharmaceuticallyacceptable excipients such as ascorbic acid, citric acid, lactic acid,acetic acid, tartaric acid, sodium sulfate, hydrochloric acid, sodiumhydroxide, sodium phosphate and sodium acetate. Glycerine can also beadded, typically to adjust the isotonicity of a parenteral formulation.

Other components can also be added to the formulations to enhance thetherapeutic effects. For example, the hydrophobic pharmaceutical agentsmay be further formulated in liposomes in addition to theabove-mentioned components. Liposomes have been shown to enhance thedelivery of compounds into cells by enhancing the compounds' ability topass through cell plasma membranes to the interior of the cells.However, because the formulations have been shown to have a therapeuticeffect with only the components described herein, formulations of thepresent invention may also “consist essentially of” or “consist of”these components.

In preferred embodiments of the invention, the formulations areeffective in treating or preventing an abnormal condition in a patientin need of such treatment. The patient is preferably a mammal and morepreferably a human. In a highly preferred embodiment, the formulationsare parenteral. Parenteral administration includes intravenous,subcutaneous, intraperitoneal, intramuscular, and intramedullaryinjection.

The term “preventing” as used herein refers to administering theformulation to a patient before the abnormal condition manifests itselfin that patient.

The term “treating” as used herein refers to the method of the inventionhaving a therapeutic effect and at least partially alleviating orabrogating the abnormal condition in the organism.

The term “therapeutic effect” as used herein refers to the inhibition ofcell growth causing or contributing to an abnormal condition. The term“therapeutic effect” also refers to the inhibition of factors causing orcontributing to the abnormal condition. A therapeutic effect relieves tosome extent one or more of the symptoms of the abnormal condition.

The term “mammal” as used herein preferably refers to such organisms asmice, rats, rabbits, guinea pigs, goats, sheep, horses, and cows, forexample; more preferably to dogs, cats, monkeys, and apes; and mostpreferably to humans.

The term “cell proliferative disorder” as used herein refers to adisorder where an excess cell proliferation of one or more subset ofcells in a multicellular organism occurs resulting in harm (e.g.,discomfort or decreased life expectancy) to the multicellular organism.The excess cell proliferation can be determined by reference to thegeneral population and/or by reference to a particular patient (e.g., atan earlier point in the patient's life). Hyper-proliferative celldisorders can occur in different types of animals and in humans, andproduce different physical manifestations depending upon the affectedcells. Hyper-proliferative cell disorders include cancers, blood vesselproliferative disorders, fibrotic disorders, and autoimmune disorders.

In reference to the treatment of abnormal cell proliferative conditions,a therapeutic effect refers to one or more of the following: (a) areduction in tumor size; (b) inhibition (i.e., slowing or stopping)tumor metastasis; (c) inhibition of tumor growth; and (d) relieving tosome extent one or more of the symptoms associated with the abnormalcondition. Compounds demonstrating efficacy against leukemias can beidentified as described herein, except that rather than inhibitingmetastasis, the compounds may instead slow or decrease cellproliferation or cell growth.

The term “abnormal condition” refers to a function in the cells ortissues of a patient that deviates from their normal functions in thatpatient. An abnormal condition can relate to cell proliferation asdescribed herein.

The present invention also features oral formulations for hydrophobicpharmaceutical agents, such as quinazoline-, nitrothiazole- andindolinone-based compounds. The oral formulations, which include one ormore polyoxyhydrocarbyl compounds, one or more polyglycolized lipids,and one or more surfactants, also have advantageous solubilitycharacteristics and oral bioavailability. These formulations allow forthe oral administration of the hydrophobic pharmaceutical agents fortesting and therapy and have shown efficacy in the preclinicalangiogenesis mice model.3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone is the most widelytested of Applicant's indolinone-based compounds. The oral formulationsof 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone have showntherapeutic effect in test animals.

Thus, a featured aspect of the invention is a formulation comprising:(a) one or more hydrophobic pharmaceutical agents, where the agents areindependently selected from the group consisting of quinazoline-,nitrothiazole- and indolinone-based compounds; (b) one or morepolyoxyhydrocarbyl compounds; (c) one or more polyglycolized lipids; and(d) one or more pharmaceutically acceptable surfactants.

It is anticipated that the one or more hydrophobic pharmaceutical agentsmay include a combination of nitrothiazole-based compounds withquinazoline-based compounds, or nitrothiazole-based compounds withindolinone-based compounds, or quinazoline-based compounds withindolinone-based compounds. In addition, the one or more hydrophobicpharmaceutical agents may include a combination of indolinone-basedcompounds, for example3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-amino)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-chloro)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, and3-[(3-methylthiophenyl-5-yl)methylene]-(4-methyl)-2-indolinone. Anotherpossiblity is that the one or more hydrophobic pharmaceutical agents mayinclude a combination of quinazoline-based compounds, for example4-(3-bromophenyl)-6,7-dimethoxyquinazoline and4-(3-chlorophenyl)-6,7-dimethoxyquinazoline. Or alternatively, the oneor more hydrophobic pharmaceutical agents may include a combination ofnitrothiazole-based compounds, for example some combination of2-methyl-5-[(5-nitrothiazol-2-yl)mercapto]-1,3,4-thiadiazole;1-benzyl-5-[(5-nitrothiazol-2-yl)mercapto]tetrazole;2-[(5-nitrothiazol-2-yl)mercapto]-5-t-butyl-1,2,4-triazole;3-[(5-nitrothiazol-2-yl)mercapto]-5-(thien-2-yl)-1,2,4-triazole;3-[(5-nitrothiazol-2-yl)mercapto]-5-phenyl-1,2,4-triazole; and4-allyl-3-hydroxy-5-[(5-nitrothiazole-2-yl)mercapto]-1,2,4-triazole.

The term “polyglycolized lipids” as used herein refers to mixtures ofmonoglycerides, diglycerides, or triglycerides and polyethyleneglycolmonoesters and diesters formed by the partial alcoholysis of vegetableoil using PEG of 200 g/mol to 2,000 g/mol or by the esterification offatty acids using PEG 200 g/mol to 2,000 g/mol and glycerols. Preferablythese include GELUCIRE® 35/10, GELUCIRE® 44/14, GELUCIRE® 46/07,GELUCIRE® 50/13, GELUCIRE® 53/10, and LABRASOL®.

In preferred embodiments of the invention, the hydrophobicpharmaceutical agent is a quinazoline-based compound of formula I, whereR₁, R₂, R₃, R₄, R₅ and R₆ are independently selected from the groupconsisting of: (i) hydrogen; (ii)saturated or unsaturated alkyl; (iii)an aryl optionally substituted with one, two, or three substituentsindependently selected from the group consisting of alkyl, alkoxy,halogen, trihalomethyl, carboxylate, nitro, and ester moieties; (iv) anamine of formula —NX₂X₃, where X₂ and X₃ are independently selected fromthe group consisting of hydrogen, saturated or unsaturated alkyl, andhomocyclic or heterocyclic ring moieties; (v) halogen or trihalomethyl;(vi) a ketone of formula —CO—X₄, where X₄ is selected from the groupconsisting of alkyl and homocyclic or heterocyclic ring moieties; (vii)a carboxylic acid of formula —(X₅)_(n)—COOH or ester of formula—(X₆)_(n)—COO—X₇, where X₅, X₆, and X₇ and are independently selectedfrom the group consisting of alkyl and homocyclic or heterocyclic ringmoieties and where n is 0 or 1; (viii) an alcohol of formula (X₈)_(n)—OHor an alkoxy moiety of formula —(X₈)_(n)—O—X₉, where X₈ and X₉ areindependently selected from the group consisting of saturated orunsaturated alkyl and homocyclic or heterocyclic ring moieties, whereinsaid ring is optionally substituted with one or more substituentsindependently selected from the group consisting of alkyl, alkoxy;halogen, trihalomethyl, carboxylate, nitro, and ester and where n is 0or 1; (ix) an amide of formula —NHCOX₁₀, where X₁₀ is selected from thegroup consisting of alkyl, hydroxyl, and homocyclic or heterocyclic ringmoieties, wherein said ring is optionally substituted with one or moresubstituents independently selected from the group consisting of alkyl,alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester; (x)—SO₂NX₁₁X₁₂, where X₁₁ and X₁₂ are selected from the group consisting ofhydrogen, alkyl, and homocyclic or heterocyclic ring moieties; (xi) ahomocyclic or heterocyclic ring moiety optionally substituted with one,two, or three substituents independently selected from the groupconsisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, nitro,and ester moieties; (xii) an aldehyde of formula —CO—H; and (xiii) asulfone of formula —SO₂—X₁₃, where X₁₃ is selected from the groupconsisting of saturated or unsaturated alkyl and homocyclic orheterocyclic ring moieties.

In other preferred embodiments of the invention, the hydrophobicpharmaceutical agent is a quinazoline-based compound of formula II,where R₁, R₂, and R₃ are selected from the group consisting of halogen,trihalomethyl, cyano, methoxy, and hydrogen. Most preferably, thequinazoline-based compound is4-(3-bromophenyl)-6,7-dimethoxyquinazoline.

In yet other preferred embodiments of the invention, the hydrophobicpharmaceutical agent is a nitrothiazole-based compound of formula IV,wherein R₁, R₂, and R₃ are independently selected from the groupconsisting of: (i) hydrogen; (ii) saturated or unsaturated alkyl; (iii)an aryl optionally substituted with one, two, or three substituentsindependently selected from the group consisting of alkyl, alkoxy,halogen, trihalomethyl, carboxylate, nitro, and ester moieties; (iv) anamine of formula —NX₂X₃, where X₂ and X₃ are independently selected fromthe group consisting of hydrogen, saturated or unsaturated alkyl, andhomocyclic or heterocyclic ring moieties; (v) halogen or trihalomethyl;(vi) a ketone of formula —CO—X₄, where X₄ is selected from the groupconsisting of alkyl and homocyclic or heterocyclic ring moieties; (vii)a carboxylic acid of formula —(X₅)_(n)—COOH or ester of formula—(X₆)_(n)—COO—X₇, where X₅, X₆, and X₇ and are independently selectedfrom the group consisting of alkyl and homocyclic or heterocyclic ringmoieties and where n is 0 or 1; (viii) an alcohol of formula (X₈)_(n)—OHor an alkoxy moiety of formula —(X₈)_(n)—O—X₉, where X₈ and X₉ areindependently selected from the group consisting of saturated orunsaturated alkyl and homocyclic or heterocyclic ring moieties, whereinsaid ring is optionally substituted with one or more substituentsindependently selected from the group consisting of alkyl, alkoxy,halogen, trihalomethyl, carboxylate, nitro, and ester and where n is 0or 1; (ix) an amide of formula —NHCOX₁₀, where X₁₀ is selected from thegroup consisting of alkyl, hydroxyl, and homocyclic or heterocyclic ringmoieties, wherein said ring is optionally substituted with one or moresubstituents independently selected from the group consisting of alkyl,alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester; (x)—SO₂NX₁₁X₁₂, where X₁₁ and X₁₂ are selected from the group consisting ofhydrogen, alkyl, and homocyclic or heterocyclic ring moieties; (xi) ahomocyclic or heterocyclic ring moiety optionally substituted with one,two, or three substituents independently selected from the groupconsisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, nitro,and ester moieties; (xii) an aldehyde of formula —CO—H; and (xiii) asulfone of formula —SO₂—X₁₃, where X₁₃ is selected from the groupconsisting of saturated or unsaturated alkyl and homocyclic orheterocyclic ring moieties.

In yet other preferred embodiments of the invention, the hydrophobicpharmaceutical agent is a nitrothiazole-based compound of formula V,wherein R₁ and R₂ are independently selected from the group consistingof: (i) hydrogen; (ii) saturated or unsaturated alkyl; (iii) an aryloptionally substituted with one, two, or three substituentsindependently selected from the group consisting of alkyl, alkoxy,halogen, trihalomethyl, carboxylate, nitro, and ester moieties; (iv) anamine of formula —NX₂X₃, where X₂ and X₃ are independently selected fromthe group consisting of hydrogen, saturated or unsaturated alkyl, andhomocyclic or heterocyclic ring moieties; (v) halogen or trihalomethyl;(vi) a ketone of formula —CO—X₄, where X₄ is selected from the groupconsisting of alkyl and homocyclic or heterocyclic ring moieties; (vii)a carboxylic acid of formula —(X₅)_(n)—COOH or ester of formula—(X₆)_(n)—COO—X₇, where X₅, X₆, and X₇ and are independently selectedfrom the group consisting of alkyl and homocyclic or heterocyclic ringmoieties and where n is 0 or 1; (viii) an alcohol of formula (X₈)_(n)—OHor an alkoxy moiety of formula —(X₈)_(n)—O—X₉, where X₈ and X₉ areindependently selected from the group consisting of saturated orunsaturated alkyl and homocyclic or heterocyclic ring moieties, whereinsaid ring is optionally substituted with one or more substituentsindependently selected from the group consisting of alkyl, alkoxy,halogen, trihalomethyl, carboxylate, nitro, and ester and where n is 0or 1; (ix) an amide of formula —NHCOX₁₀, where X₁₀ is selected from thegroup consisting of alkyl, hydroxyl, and homocyclic or heterocyclic ringmoieties, wherein said ring is optionally substituted with one or moresubstituents independently selected from the group consisting of alkyl,alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester; (x)—SO₂NX₁₁X₁₂, where X₁₁ and X₁₂ are selected from the group consisting ofhydrogen, alkyl, and homocyclic or heterocyclic ring moieties; (xi) ahomocyclic or heterocyclic ring moiety optionally substituted with one,two, or three substituents independently selected from the groupconsisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, nitro,and ester moieties; (xii) an aldehyde of formula —CO—H; and (xiii) asulfone of formula —SO₂—X₁₃, where X₁₃ is selected from the groupconsisting of saturated or unsaturated alkyl and homocyclic orheterocyclic ring moieties.

In particularly preferred embodiments of the invention, thenitrothiazole-based compound is selected from the group consisting of:2-methyl-5-[(5-nitrothiazol-2-yl)mercapto]-1,3,4-thiadole;1-benzyl-5-[(5-nitrothiazol-2-yl)mercapto]tetrazole;2-[(5-nitrothiazol-2-yl)mercapto]-5-t-butyl-1,2,4-triazole;3-[(5-nitrothiazol-2-yl)mercapto]-5-(thien-2-yl)-1,2,4-triazole;3-[(5-nitrothiazol-2-yl)mercapto]-5-phenyl-1,2,4-triazole; and4-allyl-3-hydroxy-5-[(5-nitrothiazole-2-yl)mercapto]-1,2,4-triazole.

In other preferred embodiments of the invention, the hydrophobicpharmaceutical agent is an indolinone-based compound of formula VI,where R₁, R₂, R₃, and R₄ are selected from the group consisting ofhydrogen, trihalomethyl, hydroxyl, amine, thioether, cyano, alkoxy,alkyl, amino, bromo, fluoro, chloro, iodo, mercapto, thio, cyanoamido,alkylthio, aryl, heteroaryl, carboxyl, ester, oxo, alkoxycarbonyl,alkenyl, alkoxy, nitro, alkoxyl, and amido moieties; and R₅ is anoptionally substituted aryl or heteroaryl cyclic moiety.

Preferably, the indolinone-based compound has a structure of formula VI,where R₁, R₂, R₃, and R₄ are selected from the group consisting ofhydrogen, halogen, alkyl, and alkoxy; and where R₅ is a pyrrolyl orthiophenyl moiety optionally substituted with moieties selected from thegroup consisting of hydrogen, halogen, alkyl, and alkoxy.

More preferably, the indolinone-based compound is selected from thegroup consisting of3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-amino)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-chloro)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, and3-[(3-methylthiophenyl-5-yl)methylene]-(4-methyl)-2-indolinone. Mostpreferably, the indolinone-based compound is3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone.

In some embodiments of the invention, the one or more polyoxyhydrocarbylcompounds are independently selected from the group consisting of: watersoluble carbohydrates, water soluble carbohydrate derivatives,polypeptides, water soluble polymers, water soluble mixed oxyalkylenepolymers, and the polymeric form of ethylene glycol. Preferably, the oneor more polyoxyhydrocarbyl compounds are poly(ethylene glycol) (PEG) orPEG derivatives. More preferably, PEG may vary in molecular weight fromabout 200 daltons to about 20,000 daltons.

In other embodiments of the invention, the one or more polyglycolizedlipids are mixtures of monoglycerides, diglycerides, or triglyceridesand polyethyleneglycol monoesters and diesters. Preferably, the one ormore polyglycolized lipids are selected from the group consisting of:GELUCIRE® 35/10,GELUCIRE® 44/14, GELUCIRE® 46/07,GELUCIRE®50/13,GELUCIRE® 53/10, and LABRASOL®. Most preferably, thepolyglycolized lipids are selected from the group consisting ofGELUCIRE® 44/14 and LABRASOL®.

In other embodiments of the invention, the one or more surfactants areselected independently from the group consisting of pharmaceuticallyacceptable non-ionic surfactants, polyoxyethylene castor oilderivatives, and pharmaceutically acceptable anionic surfactants. Inhighly preferred embodiments, the surfactant is CREMOPHOR EL®.

Other preferred embodiments of the invention feature formulations thatalso contain one or more pharmaceutically acceptable oils selectedindependently from the group consisting of mineral oil, vegetable oil,fractionated coconut oil, propyleneglycol monolaurate, and mixedtriglycerides with caprylic acid and capric acid. In a highly preferredembodiment, the oil is Miglyol 812.

The term “pharmaceutically acceptable oils” as used herein refers tooils such as mineral oil or vegetable oil (including safflower oil,peanut oil, and olive oil), fractionated coconut oil, propyleneglycolmonolaurate, mixed triglycerides with caprylic acid and capric acid, andthe like. Preferred embodiments of the invention feature mineral oil,vegetable oil, fractionated coconut oil, mixed triglycerides withcaprylic acid, and capric acid. A highly preferred embodiment of theinvention features Miglyol 812 (available from Huls America, USA).

In yet other preferred embodiments, when the hydrophobic pharmaceuticalagent is an indolinone-based compound substituted with one or morecarboxyl moieties, the formulation also comprises water.

In other preferred embodiments, the invention relates to formulationscomprising: (a) about 3% w/w of an indolinone-based compound of formulaVI, where R₁, R₂, R₃, and R₄ are selected from the group consisting ofhydrogen, trihalomethyl, hydroxyl, amine, thioether, cyano, alkoxy,alkyl, amino, bromo, fluoro, chloro, iodo, mercapto, thio, cyanoamido,alkylthio, aryl, heteroaryl, carboxyl, ester, oxo, alkoxycarbonyl,alkenyl, alkoxy, nitro, alkoxyl, and amido moieties; and R₅ is anoptionally substituted aryl or heteroaryl cyclic moiety; (b) about 70%w/w GELUCIRE® 44/14; (c) about 10% w/w CREMOPHOR EL®; (d) about 10% w/wMiglyol 812; and (e) about 10% w/w polyethylene glycol 600.

In other preferred embodiments, the invention relates to formulationscomprising: (a) about 3% w/w of an indolinone-based compound of formulaVI, where R₁, R₂, R₃, and R₄ are selected from the group consisting ofhydrogen, trihalomethyl, hydroxyl, amine, thioether, cyano, alkoxy,alkyl, amino, bromo, fluoro, chloro, iodo, mercapto, thio, cyanoamido,alkylthio, aryl, heteroaryl, carboxyl, ester, oxo, alkoxycarbonyl,alkenyl, alkoxy, nitro, alkoxyl, and amido moieties; and R₅ is anoptionally substituted aryl or heteroaryl cyclic moiety; (b) about 76%w/w LABRASOL”; (c) about 12% w/w CREMOPHOR EL®; and (d) about 12% w/wpolyethylene glycol 600.

In other preferred embodiments, the invention relates to formulationscomprising: (a) about 3% w/w of an indolinone-based compound of formulaVI, where R₁, R₂, R₃, and R₄ are selected from the group consisting ofhydrogen, trihalomethyl, hydroxyl, amine, thioether, cyano, alkoxy,alkyl, amino, bromo, fluoro, chloro, iodo, mercapto, thio, cyanoamido,alkylthio, aryl, heteroaryl, carboxyl, ester, oxo, alkoxycarbonyl,alkenyl, alkoxy, nitro, alkoxyl, and amido moieties; and R₅ is anoptionally substituted aryl or heteroaryl cyclic moiety; (b) about 70%w/w LABRASOL®; (c) about 10% w/w CREMOPHOR EL®; (d) about 10% w/wMiglyol 812; and (e) about 10% w/w polyethylene glycol 600.

In the most preferred embodiments, these formulations feature3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone as theindolinone-based compound.

Preferably, the formulations are effective in treating or preventing anabnormal condition in a patient, preferably a mammal, more preferably ahuman, in need of such treatment. The formulation is preferablyadministered orally. Abnormal conditions which may be treated with theseformulations include cell proliferative disorders, typically thosecharacterized by abnormal protein kinase activity. Preferably, theformulation inhibits protein kinase activity.

The compositions of the invention can also include from about 0 to about3 molar equivalents (based on the amount of the indolinone-basedcompound or other hydrophobic pharmaceutical agent in the composition)of a pharmaceutically acceptable acid or base or a mixture ofpharmaceutically acceptable acids or bases. Preferably, thepharmaceutically acceptable acid or base or mixture of pharmaceuticallyacceptable acids is present in a total amount of from about 0.2 to about2.0 molar equivalents (based on the amount of the indolinone-basedcompound or other hydrophobic pharmaceutical agent in the composition).

The solution can also include from about 0% to about 10% (by weight ofthe total solution) of water, and may also include an antioxidant (forexample, ascorbic acid, BHA (butylated hydroxyanisole), BHT (butylatedhydroxytoluene), vitamin E, vitamin E PEG 1000 succinate and the like)for chemical stability. Solutions encapsulated in a SEC may also includeglycerin for physical stability.

The compositions of this invention (e.g. solution/semisolid or solid orencapsulated solution/semisolid or solid) provide improved oralbioavailability for 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinonewhen compared to non-formulated3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone.

In another aspect, the invention relates to a method of testing thesolubility of hydrophobic pharmaceutical agents in a parenteralformulation comprising one or more hydrophobic pharmaceutical agents,that are independently selected from the group consisting ofquinazoline-, nitrothiazole-, and indolinone-based compounds; one ormore polyoxyhydrocarbyl compounds; and one or more pharmaceuticallyacceptable surfactants. The method of testing the solubility of the oneor more hydrophobic pharmaceutical agents comprises the following steps:(a) interacting the formulation with a hydrophobic support in a firstsolvent; (b) eluting the hydrophobic pharmaceutical agent from thesupport with a second solvent; and (c) comparing the amount of thehydrophobic pharmaceutical agent that elutes from the support to theamount that was added to the formulation.

The term “hydrophobic support” as used herein refers to a solid matrixthat comprises hydrocarbon moieties. The solid matrix can includereverse phase silica, cellulose, and others commonly known to thoseskilled in the art. The hydrocarbon moieties can include preferablyabout four carbon atoms, more preferably about eight carbon atoms, andmost preferably about eighteen carbon atoms. The solid matrix can becontained within a high performance liquid chromatography (HPLC) column.A pump system attached to such a column can deliver solvents to thematrix at high pressures and render high resolution of compounds elutingfrom the column.

The term “interacting” as used herein with reference to the solidsupport refers to adsorbing one or more molecules in a formulation tothe solid support. Adsorbing or binding the molecules to the solidsupport can be accomplished in different solvents, preferably thosedescribed herein by example.

The term “eluting” as used herein with reference to the solid supportrefers to the processes of desorbing (removing) a hydrophobicpharmaceutical agent such as an indolinone-, a quinazoline-, or anitrothiazole-based compound adsorbed (reversibly bound) to a solidsupport. Eluting a compound from a solid support can be accomplished bychanging the solvent to one where the hydrophobic pharmaceutical agentno longer binds to the solid support. The second solvent often has adifferent pH or a different solvent content than the solvent in whichthe hydrophobic pharmaceutical agent adsorbed to the solid support. Whena HPLC column is employed in this process, a hydrophobic pharmaceuticalagent is typically bound to the column in one solvent and then eluted bypassing another solvent through the column. The hydrophobicpharmaceutical agent flows from the column with the second solvent.

The term “comparing” as used herein in reference to the method oftesting the solubility of the hydrophobic pharmaceutical agent refers todifference in the amount of a hydrophobic pharmaceutical agent added toa formulation and the amount of the hydrophobic pharmaceutical agentthat is actually dissolved in the formulation. One may determine theamount of the hydrophobic pharmaceutical agent added to the formulation,for example, by weighing the compound before adding it to theformulation. Then one can centrifuge or filter a sample of theformulation to remove any hydrophobic pharmaceutical agent that is notdissolved in the formulation. The filtered or centrifuged formulationcan then be injected onto an HPLC column and eluted from it. Theconcentration of the hydrophobic pharmaceutical agent can then bequantified by techniques commonly known to those skilled in the art,such as by using a ultraviolet detector which measures the amount of anhydrophobic pharmaceutical agent eluted from the column by itsabsorbance. The concentration of the hydrophobic pharmaceutical agenteluted from the column can be determined from its absorbance and itsinherent extinction coefficient and/or by comparing the absorbance tothat of standard amounts of the hydrophobic pharmaceutical agent.

In preferred embodiments of the method of testing the solubility ofhydrophobic pharmaceutical agents in a parenteral formulation, theparenteral formulation further comprises one or more pharmaceuticallyacceptable alcohols.

In another aspect, the invention relates to a method of testing thesolubility of hydrophobic pharmaceutical agents in an oral formulationcomprising one or more hydrophobic pharmaceutical agents that areindependently selected from the group consisting of quinazoline-,nitrothiazole-, and indolinone-based compounds; one or morepolyoxyhydrocarbyl compounds; one or more polyglycolized lipids; and oneor more pharmaceutically acceptable surfactants. The method of testingthe solubility of the one or more hydrophobic pharmaceutical agentscomprises the following steps: (a) interacting the formulation with ahydrophobic support in a first solvent; (b) eluting the hydrophobicpharmaceutical agent from the support with a second solvent; and (c)comparing the amount of the hydrophobic pharmaceutical agent that elutesfrom the support to the amount that was added to the formulation.

In preferred embodiments of the method of testing the solubility ofhydrophobic pharmaceutical agents in an oral formulation, the parenteralformulation further comprises one or more pharmaceutically acceptableoils.

In preferred embodiments of the method of testing the solubility ofhydrophobic pharmaceutical agents, the first solvent comprises phosphateand triethylamine, and the second solvent comprises triethylamine,tetrahydrofuran, and methanol. Preferably the first solvent comprises0.35 M phosphate and 0.1% triethylamine and the second solvent comprises0.1% triethylamine, tetrahydrofuran, and methanol mixed in a 40:20:40ratio.

Most preferably, the hydrophobic pharmaceutical agent is anindolinone-based compound of formula VI.

In yet another aspect, the invention relates to a method of preparing aparenteral formulation. The method comprises the following steps: (a)dissolving one or more hydrophobic pharmaceutical agents intopolyoxyhydrocarbyl compounds to form a solution, where the agents areindependently selected from the group consisting of: a quinazoline-, anitrothiazole-, and an indolinone-based compound; (b) dissolving one ormore surfactants into the solution; and (c) filtering the solution. Insome embodiments, one or more pharmaceutically acceptable alcohols areadded to the solution.

In preferred embodiments, the invention relates to the method ofpreparing a parenteral formulation comprising the following steps: (a)dissolving 0.01 to 10 g/mL PEG-400 into water; (b) dissolving anindolinone-based compound of formula VI into the solution, where R₁, R₂,R₃, and R₄ are selected from the group consisting of hydrogen,trihalomethyl, hydroxyl, amine, thioether, cyano, alkoxy, alkyl, amino,bromo, fluoro, chloro, iodo, mercapto, thio, cyanoamido, alkylthio,aryl, heteroaryl, carboxyl, ester, oxo, alkoxycarbonyl, alkenyl, alkoxy,nitro, alkoxyl, and amido moieties; and R₅ is an optionally substitutedaryl or heteroaryl cyclic moiety; (c) adding 0.01 to 1 g/mL ethanol and0.001 to 1 g/mL benzyl alcohol to the solution; (d) dissolving 0.01 to10 g/mL ethoxylated castor oil into the solution; (e) bringing thevolume of the solution to about 100 milliliters with distilled water;and (f) filtering the solution through a 0.2 Am nylon filter.

In other preferred embodiments, the invention relates to the method ofpreparing a parenteral formulation comprising the following steps: (a)dissolving about 35 grams of PEG-400 into water; (b) dissolving about500 milligrams of an indolinone-based compound of formula VI, where R₁,R₂, R₃, and R₄ are selected from the group consisting of hydrogen,trihalomethyl, hydroxyl, amine, thioether, cyano, alkoxy, alkyl, amino,bromo, fluoro, chloro, iodo, mercapto, thio, cyanoamido, alkylthio,aryl, heteroaryl, carboxyl, ester, oxo, alkoxycarbonyl, alkenyl, alkoxy,nitro, alkoxyl, and amido moieties; and R₅ is an optionally substitutedaryl or heteroaryl cyclic moiety; (c) adding about 11.4 grams of ethanoland 2 grams of benzyl alcohol to the solution; (d) dissolving about 25grams of CREMOPHOR EL® into the solution; (e) bringing the volume of thesolution to about 100 milliliters with distilled water; and (f)filtering the solution through a 0.2 μm nylon filter.

In yet another aspect, the invention relates to a method of preparing anoral formulation comprising the following steps: (a) melting one or morepolyglycolyzed lipids;(b) dissolving one or more polyoxyhydrocarbylcompounds and one or more surfactants, into the melted polyglycolyzedlipid; (c) dissolving one or more hydrophobic pharmaceutical agent intothe solution, where the agents are independently selected from the groupconsisting of a quinazoline-, a nitrothiazole-, and an indolinone-basedcompound; and (d) filtering the solution. In some embodiments,pharmaceutically acceptable oils are also dissolved into the meltedpolyglycolyzed lipid.

In preferred embodiments of the methods of making the parenteral andoral formulations, the hydrophobic pharmaceutical agent is anindolinone-based compound of formula VI where R₁, R₂, R₃, and R₄ areselected from the group consisting of hydrogen, trihalomethyl, hydroxyl,amine, thioether, cyano, alkoxy, alkyl, amino, bromo, fluoro, chloro,iodo, mercapto, thio, cyanoamido, alkylthio, aryl, heteroaryl, carboxyl,ester, oxo, alkoxycarbonyl, alkenyl, alkoxy, nitro, alkoxyl, and amidomoieties; and R₅ is an optionally substituted aryl or heteroaryl cyclicmoiety.

Preferably, the indolinone-based compound has a structure of formula VI,where R₁, R₂, R₃, and R₄ are selected from the group consisting ofhydrogen, halogen, alkyl, and alkoxy; and where R₅ is a pyrrolyl orthiophenyl moiety optionally substituted with moieties selected from thegroup consisting of hydrogen, halogen, alkyl, and alkoxy. Morepreferably, the indolinone-based compound is selected from the groupconsisting of3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-amino)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-chloro)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, and3-[(3-methylthiophenyl-5-yl)methylene]-(4-methyl)-2-indolinone. In ahighly preferred method, the indolinone-based compound is3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone.

The methods of preparing formulations of the invention can be scaled toany volume desired. Thus, even if a method specifies that the totalvolume of the solution is 100 mL, the formulation can be prepared as a 1mL sample by proportionally decreasing each component of the formulationby a factor of 100. For example, if 10 grams of PEG-400 is required fora 100 mL volume of the formulation, then a 1 mL sample of theformulation can be prepared by adding only (10 grams)×({fraction(1/100)})=0.1 grams of PEG-400.

Dissolving the components of the formulations of the invention can beaccomplished by a variety of techniques known to those skilled in theart. These techniques include stirring techniques (manually and withmagnetic stirring systems), vortexing techniques, vibration techniques,and sonication techniques. Sonication techniques are typicallyaccomplished using a steel probe that resonates at high frequencyvibrations.

In other preferred embodiments, the invention relates to the method oftreating or preventing an abnormal condition in a patient in need ofsuch treatment. The method comprises the following steps: (a) diluting aparenteral formulation into a pharmaceutically acceptable solution, saidparenteral formulation comprising one or more hydrophobic pharmaceuticalagents, that are independently selected from the group consisting ofquinazoline-, nitrothiazole-, and indolinone-based compounds; one ormore polyoxyhydrocarbyl compounds; and one or more pharmaceuticallyacceptable surfactants; and (b) parenterally administering the dilutedformulation to the patient. In some highly preferred embodiments, theformulation further comprises one or more pharmaceutically acceptablealcohols.

Preferably, the one or more hydrophobic pharmaceutical agents are chosenfrom a group selected for their positive results in one or more in vitroassays that corresponds to the disease or to the disorder to be treated.Examples of such assays are described in section III of the DetailedDescription of the Invention.

In preferred embodiments, the pharmaceutically acceptable solution isselected from the group consisting of saline, 0.45% N saline, WFI (waterfor injection), D5W (5% dextrose in water), and D5W 0.45% N saline. Theratio of the formulation volume to the pharmaceutically acceptablesolution volume is preferably 10:1 to 1:2 (v/v), more preferably 2:1 to1:3 (v/v), and most preferably 1:1, 1:2, or 1:3 (v/v).

In other preferred embodiments, the invention features a method oftreating a patient in need of such treatment using a formulation thatcomprises: (a) 0.1 to 100 mg/mL of an indolinone-based compound offormula VI, where R₁, R₂, R₃, and R₄ are selected from the groupconsisting of hydrogen, trihalomethyl, hydroxyl, amine, thioether,cyano, alkoxy, alkyl, amino, bromo, fluoro, chloro, iodo, mercapto,thio, cyanoamido, alkylthio, aryl, heteroaryl, carboxyl, ester, oxo,alkoxycarbonyl, alkenyl, alkoxy, nitro, alkoxyl, and amido moieties; andR₅ is an optionally substituted aryl or heteroaryl cyclic moiety; (b)0.01 to 10 g/mL PEG-400; (c) 0.01 to 1 g/mL ethanol; (d) 0.001 to 1 g/mLbenzyl alcohol; and (e) 0.01 to 10 g/mL ethoxylated castor oil.

In yet other methods of treatment the formulation comprises: (a) about5.0 mg/mL of an indolinone-based compound of formula VI, where R₁, R₂,R₃, and R₄ are selected from the group consisting of hydrogen,trihalomethyl, hydroxyl, amine, thioether, cyano, alkoxy, alkyl, amino,bromo, fluoro, chloro, iodo, mercapto, thio, cyanoamido, alkylthio,aryl, heteroaryl, carboxyl, ester, oxo, alkoxycarbonyl, alkenyl, alkoxy,nitro, alkoxyl, and amido moieties; and R₅ is an optionally substitutedaryl or heteroaryl cyclic moiety; (b) about 0.35 g/mL PEG-400; (c)about0.114 g/mL ethanol; (d) about 0.02 g/mL benzyl alcohol; and (e) about0.25 g/mL CREMOPHOR EL®.

In a highly preferred embodiment of a method of treatment, theformulation comprises: (a) about 4.5 mg/mL of an indolinone-basedcompound, 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone; (b) about45% w/v PEG-400; (c) about 31.5% w/v CREMOPHOR EL®; (d) about 2% w/vbenzyl alcohol; and (e) about 9.5% w/v ethanol.

In another aspect, the invention relates to a method of preventing ortreating an abnormal condition in a patient in need of treatmentcomprising the following steps: (a) preparing an acceptablepharmaceutical composition from an oral formulation, said oralformulation comprising: a hydrophobic pharmaceutical agent, one or morepolyoxyhydrocarbyl compounds, one or more polyglycolized lipids, and oneor more pharmaceutically acceptable surfactants; and (b) administeringsaid composition to said patient. Preferably, the formulation is orallyadministered. In highly preferable embodiments, the oral formulationfurther comprises one or more pharmaceutically acceptable oils.

The pharmaceutically acceptable composition is preferably selected fromthe group comprising: said oral formulation, a hard gelatin capsulefilled with said oral formulation, a soft gelatin capsule filled withsaid oral formulation, and a hard gelatin capsule filled with said oralcomposition admixed with a granulating agent to form a dry solidcomposition. In preferred embodiments, the solution is encapsulated in asoft elastic gelatin capsule (SEC) or a hard gelatin capsule.

A solid composition of the formulation can be prepared by mixing theformulation in a liquefied state with a pharmaceutically acceptablegranulating agent or a mixture of pharmaceutically acceptablegranulating agents (for example silicon dioxide, microcrystallinecellulose, starch, calcium carbonate, pectin, crospovidone, polyplasdoneand the like).

In preferred embodiments of the method of treatment, the formulationcomprises: (a) about 3% w/w of an indolinone-based compound of formulaVI, where R₁, R₂, R₃, and R₄ are selected from the group consisting ofhydrogen, trihalomethyl, hydroxyl, amine, thioether, cyano, alkoxy,alkyl, amino, bromo, fluoro, chloro, iodo, mercapto, thio, cyanoamido,alkylthio, aryl, heteroaryl, carboxyl, ester, oxo, alkoxycarbonyl,alkenyl, alkoxy, nitro, alkoxyl, and amido moieties; and R₅ is anoptionally substituted aryl or heteroaryl cyclic moiety; (b) about 70%w/w GELUCIRE® 44/14; (c) about 10% w/w CREMOPHOR EL®; (d) about 10% w/wMiglyol 812; and (e) about 10% w/w polyethylene glycol 600.

In other preferred embodiments of the method of treatment, theformulation comprises: (a) about 3% w/w of an indolinone-based compoundof formula VI, where R₁, R₂, R₃, and R₄ are selected from the groupconsisting of hydrogen, trihalomethyl, hydroxyl, amine, thioether,cyano, alkoxy, alkyl, amino, bromo, fluoro, chloro, iodo, mercapto,thio, cyanoamido, alkylthio, aryl, heteroaryl, carboxyl, ester, oxo,alkoxycarbonyl, alkenyl, alkoxy, nitro, alkoxyl, and amido moieties; andR₅ is an optionally substituted aryl or heteroaryl cyclic moiety; (b)about 76% w/w LABRASOL®; (c) about 12% w/w CREMOPHOR EL®; and (d) about12% w/w polyethylene glycol 600.

In yet other preferred embodiments of the method of treatment, theformulation comprises: (a) about 3% w/w of an indolinone-based compoundof formula VI, where R₁, R₂, R₃, and R₄ are selected from the groupconsisting of hydrogen, trihalomethyl, hydroxyl, amine, thioether,cyano, alkoxy, alkyl, amino, bromo, fluoro, chloro, iodo, mercapto,thio, cyanoamido, alkylthio, aryl, heteroaryl, carboxyl, ester, oxo,alkoxycarbonyl, alkenyl, alkoxy, nitro, alkoxyl, and amido moieties; andR₅ is an optionally substituted aryl or heteroaryl cyclic moiety; (b)about 70% w/w LABRASOL®; (c) about 109 w/w CREMOPHOR EL®; (d) about 10%w/w Miglyol 812; and (e) about 10% w/w polyethylene glycol 600.

In preferred embodiments of the method of treatment, the parenteral ororal formulations comprise an indolinone-based compound having astructure of formula VI, where R₁, R₂, R₃, and R₄ are selected from thegroup consisting of hydrogen, trihalomethyl, hydroxyl, amine, thioether,cyano, alkoxy, alkyl, amino, bromo, fluoro, chloro, iodo, mercapto,thio, cyanoamido, alkylthio, aryl, heteroaryl, carboxyl, ester, oxo,alkoxycarbonyl, alkenyl, alkoxy, nitro, alkoxyl, and amido moieties; andR₅ is an optionally substituted aryl or heteroaryl cyclic moiety.

In other preferred embodiments of the method of treatment, theparenteral or oral formulations comprise an indolinone-based compoundhaving a structure of formula VI, where R₁, R₂, R₃, and R₄ are selectedfrom the group consisting of hydrogen, halogen, alkyl, and alkoxy; andwhere R₅ is a pyrrolyl or thiophenyl moiety optionally substituted withmoieties selected from the group consisting of hydrogen, halogen, alkyl,and alkoxy.

In other preferred embodiments of the method of treatment, theparenteral or oral formulations comprise an indolinone-based compoundselected from the group consisting of3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-amino)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-chloro)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, and3-[(3-methylthiophenyl-5-yl)methylene]-(4-methyl)-2-indolinone.

In highly preferred embodiments of the method of treatment, theparenteral or oral formulations comprise the indolinone-based compound,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone.

In highly preferred embodiments of the invention, the method oftreatment is effective in treating or preventing an abnormal conditionin a patient, preferably a mammal, more preferably a human, in need ofsuch treatment. Abnormal conditions which may be treated with theseformulations are cell proliferative disorders, typically thosecharacterized by abnormal protein kinase activity. Preferably, theformulation inhibits protein kinase activity.

Other features and advantages of the invention will be apparent from thefollowing description of the preferred embodiments and from the claims.

DESCRIPTION OF FIGURES

FIGS. 1A and B shows a summary of exemplary formulations tested for oralbioavailability.

FIGS. 2A and B shows a summary of exemplary dosing regimens for the oralbioavailability studies.

DETAILED DESCRIPTION OF THE INVENTION

The present invention features parenteral and oral formulations forsolubilizing hydrophobic pharmaceutical agents, including quinazoline-,nitrothiazole- and indolinone-based compounds. The formulations can beused to facilitate administration of hydrophobic pharmaceuticalcompounds to patients in need of such treatment.

The invention is directed in part towards administering hydrophobicpharmaceutical agents in parenteral and oral formulations thatobliterate tumors by severing their sources of sustenance. Thehydrophobic pharmaceutical agents are designed to specifically bindprotein kinases over-expressed in the vasculature that supply tumorswith sustenance. One such protein kinase target (particularly ofindolinone-based compounds) is FLK-1, which is over-expressed in theproliferating endothelial cells of a growing tumor, but not in thesurrounding quiescent endothelial cells (Plate et al., 1992, Nature359:845-848).

FLK-1 is activated upon binding VEGF, a strong regulator for endothelialcell proliferation as well as normal and pathological angiogenesis(Klagsburn and Soker, 1993, Current Biology 3:699-702). Thus, compoundsthat specifically inhibit the FLK protein kinase are potentialanti-cancer agents as they may decrease the vasculature that nourishestumors. These inhibitors will most likely result in minimizing and evenobliterating solid tumors. In addition, compounds that specificallyinhibit FLK will potentially represent a new generation of cancertherapeutics as they will most likely cause few side effects. Thesepotential properties are a welcome improvement over the currentlyutilized cancer therapeutics that cause multiple side effects anddeleteriously weaken patients.

Another kinase target of the hydrophobic pharmaceutical agents of theinvention (particularly quinazoline-based compounds) is RAF, aserine/threonine protein kinase. RAF is a non-receptor protein kinasethat is recruited to the cell membrane when it binds to activated RAS, aguanine triphosphate hydrolyzing enzyme. RAS is activated when anactivated receptor protein tyrosine kinase, such as EGFR or PDGFR, bindto an adaptor protein, GRB2, and a guanine nucleotide exchange factor,SOS. SOS removes guanine diphosphate from RAS, replaces it with guaninetriphosphate, and thereby activates RAS. RAS then binds RAF andconsequently activates RAF. RAF may then phosphorylate other proteintargets on serine and threonine residues, such as the kinase (MEK) thatphosphorylates and consequently activates mitogen-activated proteinkinase (MAPK). Thus, RAF serves as an intermediary controlling factor inmitogen-activated signal transduction.

Due to the important regulatory role of RAF in cells, modifications tothe amino acid sequence of RAF can alter its function and consequentlymodify cellular behavior. RAF's role in cell proliferation isunderscored by the observation that mutations to RAF's amino acidsequence have been associated with tumors and cancers. Because themutations to RAF that give rise to cancer in cells lead to RAF moleculesthat display unregulated catalytic activity, inhibitors of RAF mayalleviate or even abrogate the cell proliferation that leads to cancerin these cells.

Some quinazoline-based compounds are known to inhibit the function ofthe RAF protein kinase (U.S. application Ser. No. 60/045,351, by Tang etal., filed May 2, 1997). Because RAF exhibits significant amino acidhomology to other serine/threonine protein kinases, quinazoline-basedcompounds may inhibit serine/threonine protein kinases other than RAF.

Yet another target of the hydrophobic pharmaceutical agents of thepresent invention (and in particular nitrothiazole-based compounds) areprotein tyrosine phosphatases (PTPs). Protein tyrosine phosphatasescomprise a family of transmembrane and cytoplasmic enzymes that areinvolved in cell signaling cascades. The substrates of PTPs may beprotein tyrosine kinases (PTKs) which possess phosphotyrosine residuesor the substrates of PTKs (Hunter, 1989, Cell 58:1013-16; Fischer etal., 1991, Science 253:401-6; Saito & Streuli, 1991, Cell Growth andDifferentiation 2:59-65; Pot and Dixon, 1992, Biochem. Biophys. Acta,1136:35-43). A common mechanism by which receptors regulate cellfunction is through an inducible tyrosine kinase activity which iseither endogenous to the receptor or is imparted by other proteins thatbecome associated with the receptor (Darnell et al., 1994, Science264:1415-1421; Heldin, 1995, Cell 80:213-223; Pawson, 1995, Nature373:573-580).

Protein tyrosine kinases comprise a large family of transmembranereceptor and intracellular enzymes with multiple functional domains(Taylor et al., 1992 Ann. Rev. Cell Biol. 8:429-62). Included among thePTKs are epidermal growth factor receptor (EGFR) and platelet-derivedgrowth factor receptor (PDGFR), which undergo oligomerization uponligand binding, and the receptors self-phosphorylate (viaautophosphorylation or transphosphorylation) on specific tyrosineresidues in the cytoplasmic portions of the receptor (Schlessinger andUllrich, 1992, Neuron, 9:383-91, Heldin, 1995, Cell 80:213-223). Othermembers include cytoplasmic protein tyrosine kinases (CPTKs), such asJanus kinases (e.g., JAK1, JAK2, TYK2), Src kinases (e.g., src, lck,fyn) that are associated with receptors for cytokines (e.g., IL-2, IL-3,IL-6, erythropoietin) and interferons, and antigen receptors. Thesereceptors also undergo oligomerization, and have tyrosine residues thatbecome phosphorylated during activation, but the receptor polypeptidesthemselves do not possess kinase activity.

The levels of tyrosine phosphorylation required for normal cell growthand differentiation at any time are achieved through the coordinatedaction of PTKs and PTPs. Depending on the cellular context, these twotypes of enzymes may either antagonize or cooperate with each otherduring signal transduction. An imbalance between these enzymes mayimpair normal cell functions leading to metabolic disorders and cellulartransformation.

I. Target Diseases to be Treated by Hydrophobic Pharmaceutical AgentFormulations

The hydrophobic pharmaceutical agents of the invention can be used askinase inhibitors, antimetastatic or anticancer agents, or to controlangiogenesis; for inhibiting atheromatous plaque development; fortreating Alzheimer's disease; and as immunomodulators. The currentinvention can be used in the treatment of psoriasis, epidermalhyperproliferation, restenosis, diabetic complications, and asimmunosuppressants.

Protein kinases are essential regulatory molecules that control avariety of cellular functions. For this reason, any alteration in thefunction of a protein kinase can cause an abnormal condition in anorganism. One of the many functions controlled by protein kinases iscell proliferation.

Alterations in the function of a protein kinase that normally regulatescell proliferation can lead to enhanced or decreased cell proliferativeconditions evident in certain diseases. Aberrant cell proliferativeconditions include cancers such as fibrotic and mesangial disorders,abnormal angiogenesis and vasculogenesis, wound healing, psoriasis,restenosis, diabetes mellitus, and inflammation.

Fibrotic disorders relate to the abnormal formation of the cellularextracellular matrix. An example of a fibrotic disorder is hepaticcirrhosis. Hepatic cirrhosis is characterized by an increasedconcentration of extracellular matrix constituents resulting in theformation of a hepatic scar. Hepatic cirrhosis can cause diseases suchas cirrhosis of the liver.

Mesangial cell proliferative disorders occur due to the abnormalproliferation of mesangial cells. Mesangial proliferative disordersinclude various human renal diseases, such as glomerulonephritis,diabetic nephropathy, malignant nephrosclerosis, thromboticmicroangiopathy syndromes, transplant rejection, and glomerulopathies.

Angiogenic and vasculogenic disorders result from excess proliferationof blood vessels. Blood vessel proliferation is necessary in a varietyof normal physiological processes such as embryonic development, corpusluteum formation, wound healing and organ regeneration. However, bloodvessel proliferation is also essential in cancer tumor development.Other examples of blood vessel proliferative disorders includearthritis, where new capillary blood vessels invade the joint anddestroy cartilage. In addition, blood vessel proliferative diseasesinclude ocular diseases, such as diabetic retinopathy, where newcapillaries in the retina invade the vitreous, bleed and causeblindness. Conversely, disorders related to the shrinkage, contractionor closing of blood vessels, such as restenosis, are also implicated inadverse regulation of protein kinases and protein phosphatases.

Moreover, vasculogenesis and angiogenesis are associated with the growthof malignant solid tumors and metastasis. A vigorously growing cancertumor requires a nutrient and oxygen rich blood supply to continuegrowing. As a consequence, an abnormally large number of capillary bloodvessels often grow in concert with the tumor and act as supply lines tothe tumor. In addition to supplying nutrients to the tumor, the newblood vessels embedded in a tumor provide a gateway for tumor cells toenter the circulation and metastasize to distant sites in the organism(Folkman, 1990, J. Natl. Cancer Inst. 82:4-6).

II. Synthesis of Hydrophobic Pharmaceutical Agents

The indolinone-based compounds of the invention are synthesized byreacting an aldehyde with an indolinone. Descriptions of methods forsynthesizing indolinone-based compounds are provided in U.S. applicationSer. No. 08/702,282 and PCT Publication No. WO 96/40116, published Dec.19, 1996, both of which are incorporated herein by reference in theirentirety, including any drawings and figures. The examples fullydescribe the solvents, temperatures, separation techniques, and otherconditions utilized for the invention. Other synthetic techniques, suchas those described in International patent publication WO 96/22976,published Aug. 1, 1996 may also be used or modified by those skilled inthe art to make the compounds of the resent invention.

The quinazoline-based compounds of the invention are synthesizedfollowing the description of the methods provided in U.S. applicationSer. No. 60/045,351, by Tang et al., filed May 2, 1997, incorporatedherein by reference in its entirety, including any drawings or figures.The examples fully describe the solvents, temperatures, separationtechniques, and other conditions utilized for the invention.

The quinazoline compounds of the invention are synthesized using thefollowing general procedure unless otherwise stated:

(i) evaporation were carried out by rotary evaporation in vacuo;

(ii) operations were carried out under an atmosphere of an inert gassuch as nitrogen;

(iii) high performance liquid chromatography (HPLC) were performed onMerck LiChrosorb RP-18 reversed-phase silica obtained from E. Merck,Darmstadt, Germany;

(iv) yields are given for illustration only and are not necessarily themaximium attainable;

(v) melting points are uncorrected and were determined using a HWS MainzSG 2000 digital melting point apparatus;

(vi) the structures of all compounds of the formula (I), (II), and (III)of this invention were confirmed by proton magnetic resonancespectroscopy on a Bruker AMX500-NMR spectrophotometer, by elementalmicroanalysis and, in certain cases, by mass spectroscopy;

(vii) the purity of the structures were performed by thin layerchromatography (TLC) using silica gel (Merck Silica Gel 60 F254) or byHPLC;

(viii) intermediates were not generally fully characterised and puritywas assessed by thin layer chromatography (TLC) or by HPLC.

It should be understood that the invention is not limited to theparticular embodiments shown and described herein, but that variouschanges and modifications may be made without departing from the spiritand scope of the invention as defined by the claims. The followingspecific procedures were utilized to synthesize quinazoline compounds ofthe invention.

Procedure A

Method for Reaction of 2,4-Diamino-5-Fluoroquinazoline with SodiumPhenolates and Thiophenolates:

Dimethyl sulphoxide and sodium hydride (80% disp in mineral oil) wasadded to a dry flask maintained under inert atmosphere at roomtemperature. A solution of phenol (optionally substituted) in dimethylsulphoxide was added to the stirred reaction mixture, heated to 60° C.for 30 minutes and allowed to cool. 2,4-Diamino-5-fluoroquinazoline wasadded all at once (as the solid) and the reaction mixture was heated toca 150° C. for 2-3 hours. After cooling to room temperature thesuspension was diluted with water and e.g. with methanol, the solidcollected by filtration, washed, recrystallized and dried at 50° C. invacuo. 2,4-Diamino-5-fluoroquinazoline was prepared from2,6-difluorobenzonitrile (Lancaster, Acros) according to the publishedmethod (J. Heterocyclic. Chem. 25, 1173 (1988)).

The following compounds have been obtained according to this procedure:

1. 2,4-Diamino-5-(4-methoxyphenoxy)quinazoline, m.p. 268-270° C.

2. 2,4-Diamino-5-(3-trifluormethylphenoxy)quinazoline, m.p. 280-284° C.(dec)

 2,4-Diamino-5-phenylthioquinazoline was also synthsized by thefollowing method: 2,4-Diamino-5-fluoroquinazoline (3.6 g, 20 mmol) andsodium thiophenolate (Fluka) (3.2 g, 24 mmol) in DMSO (100 ml) werereacted at 75° C. for 15 hours. After cooling to room temperature thesuspension was diluted with water (25 ml) and methanol (150 ml), thesolid was collected by filtration, washed with methanol, recrystallizedfrom n-butyl acetate and dried at 50° C. in vacuo to give2,4-diamino-5-phenylthioquinazoline (1.0 g, 18.60%, m.p. 240-244° C.):

Procedure B

Method for Reaction of 2,4-Diamino-5-Fluoroquinazoline with PotassiumPhenolates:

To a stirred solution of potassium tert-butoxide in dimethyl sulphoxideunder nitrogen phenol (optionally substituted) and after evolution ofhydrogen had ceased 2,4-diamino-5-fluoroquinazoline were added all atonce (as the solid) and the mixture was heated to ca 150° C. for 2-3hours. After cooling to room temperature the suspension was diluted withwater and e.g. with methanol, the solid was collected by filtration,washed, and dried at 50° C. in vacuo.

The following compounds have been obtained according to this procedure:

1. 2,4-Diamino-5-(4-tert-butylphenoxy)quinazoline, m.p. 226-228° C.

2. 2,4-Diamino-5-(3,4-dimethoxyphenoxy)quinazoline, m.p. 301-302° C.

3. 2,4-Diamino-5-(3-dimethylaminophenoxy)quinazoline, m.p. 224-225° C.(dec)

4. 2,4-Diamino-5-(2-fluorophenoxv)quinazoline, m.p. 301∝303° C.

5. 2,4-Diamino-5-(3-bromophenoxy)quinazoline, m.p. 292-295° C.

6. 2,4-Diamino-5-(2-methoxyphenoxy)quinazoline, m.p. 208-209° C. (dec)

7. 2.4-Diamino-5-(3-methoxyphenoxy)quinazoline, m.p. 215-216° C. (dec)

8. 2,4-Diamino-5-(4-benzyloxyphenoxy)quinazoline, m.p. 175-177° C.

9. 2,4-Diamino-5-(3-dimethylaminopropoxy)quinazoline, m.p. 193-195° C.

In addition, the following compounds may be synthesized by either methodA or method C, described below:

Procedure C

Method for Reaction of 4-Amino-5-Fluoroquinazoline with PotassiumPhenolates and Sodium Thiophenolate:

A solution of phenol (optionally substituted) in dimethyl suiphoxide wasadded to a stirred mixture of potassium tert-butoxide in dimethylsuiphoxide at room temperature. After 15 minutes2,4-diamino-5-fluoroquinazoline was added all at once (as the solid) andthe mixture was heated to ca 50° C. for 7 hours. After cooling to roomtemperature the suspension was diluted with water, the solid collectedby is filtration, washed, recrystallized from ethanol or n-butyl acetateand dried at 50° C. in vacuo. In particular, 4-Amino-5-fluoroquinazolinewas prepared from 2,6-difluorobenzonitrile (Lancaster, Acros) accordingto the published method (J. Heterocyclic. Chem. 28, 1357 (1991)).

The following compounds have been obtained according to this procedure:

1. 4-Amino-5-(4-methoxyphenoxy)quinazoline, m.p. 192-195° C.

2. 4-Amino-5-(3-dimethylaminophenoxy)quinazoline, m.p. 179-181° C.

3. 4-Amino-5-(3-pyridinoxy)quinazoline, m.p. 245-247° C.

4. 4-Amino-5-(4-benzyloxyphenoxy)quinazoline, m.p. 170-171° C.

5. 4-Amino-5-(3,4-methylenedioxyphenoxy)quinazoline, m.p. 201-203° C.

Procedure D

Method for Reaction of 6-Substituted 2-Fluorobenzonitriles withGuanidine Carbonate:

A mixture of 1 equivalent of 6-substituted 2-fluorobenzonitriles(Maybridge, Lancaster) and 1.5 equivalent of guanidine carbonate inN,N-dimethylacetamide was heated under nitrogen at 140-150° C. for 5-6hours. The reaction mixture was allowed to cool to room temperatureovernight. The resulting suspension was diluted with water and e.g. withmethanol, and after cooling to 4° C. the solid was collected byfiltration, washed, recrystallized from n-butyl acetate, filtrated againand dried at 50° C. in vacuo.

The following compounds have been obtained according to this procedure:

1. 2,4-Diamino-5-(4-chlorophenylthio)quinazoline, m.p. 220-224° C.

2. 2,4-Diamino-5-(4-methylphenylthio)quinazoline, m.p. 206-207° C.

3. 2,4-Diamino-5-methoxyquinazoline, m.p. 199-202° C.

4. 2,4-Diamino-5-(pyrrol-1-yl)quinazoline, m.p. 248-250° C.

In similar synthetic processes, the following compounds can besynthesized:

1. 4-Amino-5-(4-fluorophenoxy)quinazoline was prepared from2-fluoro-6-(4-fluorophenoxy) benzonitrile (Maybridge) as follows:

A mixture of 2-fluoro-6-(4-fluorophenoxy)benzonitrile (2.5 g, 11 mmol)and formamidine acetate (Aldrich) (2.3 g, 22 mmol) in 50 ml ofN,N-dimethylacetamide was heated at 162° C. under nitrogen for 9 hours.After cooling to room temperature the reaction mixture was evaporatedunder reduced pressure. The product was suspended in 80 ml of cold waterand the pH adjusted to 8.5 with concentrated ammonium hydroxide. Aftercooling the suspension overnight, the precipitate was isolated byfiltration, washed with water (25 ml), dried and recrystallized from 30ml of ethanol at 4° C. The precipitate was collected by filtration,washed with ethanol and dried at 50° C. in vacuo to give4-amino-5-(4-fluorophenoxy)quinazoline (0.3 g, 10.7%, m.p. 188-190° C.):

2. 4-Amino-2-phenyl-5-phenoxyquinazoline was prepared from2-fluoro-6-phenoxybenzonitrile (Maybridge) as follows:

A mixture of 2-fluoro-6-phenoxybenzonitrile (2.7 g, 13 mmol),benzamidine hydrochloride (Aldrich) (3.0 g, 19 mmol) and sodium acetate(1.6 g, 19 mmol) in 60 ml of N,N-dimethylacetamide was heated at 150° C.under nitrogen for 6.5 hours. After cooling to room temperature thereaction mixture was evaporated under reduced pressure. The product wassuspended in 20 ml of ethanol and water (100 ml) and concentratedammonium hydroxide (10 ml) was added. The precipitate was isolated byfiltration, washed with water, dried and recrystallized twice from 60 mlof 2-propanol at 4° C. The precipitate was collected by filtration,washed with 2-propanol and dried at 50° C. in vacuo to give4-amino-2-phenyl-5-phenoxyquinazoline (0.5 g, 12%, m.p. 190-191° C.):

The following procedures can be utilized to synthesize compounds relatedto those described herein:

Procedure E

Method for Reaction of 5-Substituted 4-Aminoquinazolines with ArylIsocyanates:

To a stirred solution of the 5-substituted 4-aminoquinazoline indichloromethane aryl isocyanate (optionally substituted) was added atroom temperature and stirring continued overnight. The precipitate wascollected, washed with dichloromethane and dried at 50° C. in vacuo.

1-[5-(4-Methoxyphenoxy)Quinazolin-4-Yl]-3-Phenylurea:

4-Amino-5-(4-methoxyphenoxy)Quinazoline (1.0 g, 3.7 mmol) and phenylisocyanate (0.52 g, 4.4 mmol) were reacted in 30 ml of dichloromethaneaccording to procedure E to give1-[5-(4-methoxyphenoxy)quinazolin-4-yl]-3-phenylurea (0.9 g, 64.3%, m.p.231-232° C.).

1-[5-(4-Methoxyphenoxy)Quinazolin-4-Yl]-3-(3-Bromophenyl)Urea:

4-Amino-5-(4-methoxyphenoxy)quinazoline (0.35 g, 1.3 mmol) and3-bromophenyl isocyanate (0.31 g, 1.6 mmol) were reacted in 20 ml ofdichloromethane according to procedure E to give1-[5-(4-methoxyphenoxy)quinazolin-4-yl]-3-(3-bromophenyl)urea (0.5 g,83.3%, m.p. 249-251° C.)

1-[5-(4-Methoxyphenoxy)Quinazolin-4-Yl]-3-(3-Methoxyphenyl)Urea:

4-Amino-5-(4-methoxyphenoxy)quinazoline (0.35 g, 1.3 mmol) and3-methoxyphenyl isocyanate (0.23 g, 1.6 mmol) were reacted in 20 ml ofdichloromethane according to procedure E to give1-[5-(4-methoxyphenoxy)quinazolin-4-yl]-3-(3-methoxyphenyl)urea (0.4 g,74.1%, m. p. 209-210° C.).

4-Amino-5-Phenylthioquinazoline

4-Amino-5-fluoroquinazoline (3.2 g, 20 mmol) and sodium thiophenolate(Fluka) (4.0 g, 30 mmol) in DMSO (80 ml) were reacted at 150° C. for 5hours. After cooling to room temperature the suspension was diluted withwater (100 ml) and ethanol (50 ml), the solid was collected byfiltration, washed (water/ethanol 1:1), recrystallized from 2-propanoland dried at 50° C. in vacuo to give 2,4-diamino-5-phenylthioquinazoline(2.1 g, 41.4%, m.p. 195-197° C.).

2,4-Diamino-5-Anilinoquinazoline

Aniline (Aldrich) (5 g, 50 mmol), sodium hydride (80% disp in mineraloil) (1.5 g, 50 mmol) and 2,4-diamino-5-fluoroquinazoline (4.4 g, 25mmol) were reacted in 80 ml of dimethyl sulphoxide according toprocedure A to give 2,4-diamino-5-anilinoquinazoline (0.3 g, 4.8%, m.p.279-283° C. (dec)).

4-Acetamido-5-(4-Methoxyphenoxy)Quinazoline was Prepared from4-amino-5-(4Methoxyphenoxy)Quinazoline as follows:

To a stirred solution of 4-amino-5-(4-methoxyphenoxy)quinazoline (1.0 g,3.7 mmol) in 30 ml of dichloromethane pyridine (0.3 g, 3.7 mmol) andacetic anhydride (0.38 g, 3.7 mmol) were added at room temperature andstirring continued for 4 days. After evaporation under reduced pressure,30 ml of 2-propanol was added and after cooling to 4° C. the solid wascollected by filtration, washed, recrystallized from ethanol, filtratedagain and dried at 50° C. in vacuo. to give4-acetamido-5-(4methoxyphenoxy)quinazoline (0.5 g, 45.4%, m.p. 174-175°C.).

4-Amino-5-(4-Hydroxyphenoxy)Quinazoline

4-Amino-5-(4-benzyloxyphenoxy)quinazoline (1.5 g, 4.4 mmol) washydrogenated under 5 atm of H₂ in the presence of 0.5 g of 10% Pd/C in80 ml of N,N-dimethylacetamide at 50-60° C. After 4 hours the reactionmixture was filtered through a glass filter of silica gel, concentrated,dissolved in 80 ml of ethanol/water 4:1 (v/v) and crystallized at 4° C.The precipitate was collected by filtration, washed with ethanol anddried at 50° C. in vacuo to give 4-amino-5-(4-hydroxyphenoxy)quinazoline(0.3 g, 27.3%, m.p. 300-302° C. (dec)).

2,4-Diamino-5-(4-Hydroxyphenoxy)Quinazoline

2,4-Diamino-5-(4-benzyloxyphenoxy)quinazoline (3.6 g, 10 mmol) washydrogenated under 4 atm of H₂ in the presence of 0.36 g of 10% Pd/C in80 ml of N,N-dimethylacetamide at 50-60° C. After 4 hours the reactionmixture was filtered through a glass filter of silica gel, concentrated,dissolved in 50 ml of 2-propanol and crystallized at 4° C. Theprecipitate was collected by filtration, washed with 2-propanol anddried at 50° C. in vacuo to give2,4-diamino-5-(4-hydroxyphenoxy)quinazoline (2.3 g, 85.2%, m.p. 330-341°C. (dec)).

The nitrothiazole-based compounds of the invention are synthesized withreadily available materials using standard organic synthetic chemistrytechniques in accordance with the teachings of U.S. Pat. Nos. 5,198,333,3,970,725, and 3,850,939 which are hereby incorporated by referenceherein, including any drawings or figures.

III. Biological Activity of Hydrophobic Pharmaceutical Agents

Indolinone-based compounds of the invention have been tested for theirability to activate or inhibit protein kinases in biological assays. Themethods used to measure indolinone-based compound modulation of proteinkinase function are described in U.S. application Ser. No. 08/702,282,filed Aug. 23, 1996, incorporated herein by reference in its entirety,including any drawings. In particular, indolinone-based compounds of theinvention were tested for their ability to inhibit the FLK proteinkinase. The preferred indolinone-based compound of the invention,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, is a specific andpotent inhibitor of VEGF-induced endothelial cell growth.

Quinazoline-based compounds of the present invention have been testedfor their ability to inhibit RAF protein kinase function. The biologicalassays and results of these inhibition studies are described in U.S.application Ser. No. 60/045,351, by Tang et al., filed May 2, 1997. Themethods used to measure quinazoline-based compound modulation of proteinkinase function are similar to those described in U.S. application Ser.No. 08/702,282, filed Aug. 23, 1996, with respect to the high throughputaspect of the method. The Ser. No. 08/702,282 application isincorporated herein by reference in its entirety, including anydrawings.

Nitrothiazole-based compounds of the present invention have been testedfor their ability to inhibit protein tyrosine phosphatases by variousprocedures known in the art. Biological and biochemical assays andresults of these inhibition studies are described in U.S. applicationSer. No. 08/660,900, by Tang et al., filed Jun. 7, 1996, which is herebyincorporated herein in its entirety, including any drawings or figures.In general, such assays involve exposing target cells in culture to thecompounds and a) biochemically analyzing cell lysates to assess thelevel and/or identity of tyrosine phosphorylated proteins; or (b)scoring phenotypic or functional changes in treated cells as compared tocontrol cells that were not exposed to the test substance.

IV. Administration of Hydrophobic Pharmaceutical Agent

Formulations

Methods of determining the dosages of compounds to be administered to apatient and modes of administering compounds to an organism aredisclosed in U.S. application Ser. No. 08/702,282, filed Aug. 23, 1996and International patent publication number WO 96/22976, publishedAugust 1 1996, both of which are incorporated herein by reference in itsentirety, including any drawings. Those skilled in the art willappreciate that such descriptions are applicable to the presentinvention and can be easily adapted to it.

The proper dosage depends on various factors such as the type of diseasebeing treated, the particular composition being used, and the size andphysiological condition of the patient. Therapeutically effective dosesfor the compounds described herein can be estimated initially from cellculture and animal models. For example, a dose can be formulated inanimal models to achieve a circulating concentration range thatinitially takes into account the IC₅₀ as determined in cell cultureassays. The animal model data can be used to more accurately determineuseful doses in humans.

Plasma half-life and biodistribution of the drug and metabolites in theplasma, tumors, and major organs can be also be determined to facilitatethe selection of drugs most appropriate to inhibit a disorder. Suchmeasurements can be carried out. For example, HPLC analysis can beperformed on the plasma of animals treated with the drug and thelocation of radiolabeled compounds can be determined using detectionmethods such as X-ray, CAT scan, and MRI. Compounds that show potentinhibitory activity in the screening assays, but have poorpharmacokinetic characteristics, can be optimized by altering thechemical structure and retesting. In this regard, compounds displayinggood pharmacokinetic characteristics can be used as a model.

Toxicity studies can also be carried out by measuring the blood cellcomposition. For example, toxicity studies can be carried out in asuitable animal test (e.g., mice in the example below) as follows: 1)the compound is administered to mice (an untreated control mouse shouldalso be used); 2) blood samples are periodically obtained via the tailvein from one mouse in each treatment group; and 3) the samples areanalyzed for red and white blood cell counts, blood cell composition,and the percent of lymphocytes versus polymorphonuclear cells. Acomparison of results for each dosing regime with the controls indicatesif toxicity is present.

At the termination of each toxicity study, further studies can becarried out by sacrificing the animals (preferably, in accordance withthe American Veterinary Medical Association guidelines Report of theAmerican Veterinary Medical Assoc. Panel on Euthanasia, Journal ofAmerican Veterinary Medical Assoc., 202:229-249, 1993). Representativeanimals from each treatment group can then be examined by gross necropsyfor immediate evidence of metastasis, unusual illness, or toxicity.Gross abnormalities in tissue are noted, and tissues are examinedhistologically. Compounds causing a reduction in body weight or bloodcomponents are less preferred, as are compounds having an adverse effecton major organs. In general, the greater the adverse effect the lesspreferred the compound.

For the treatment of cancers the expected daily dose of a hydrophobicpharmaceutical agent is between 1 to 500 mg/day, preferably 1 to 250mg/day, and most preferably 1 to 50 mg/day. Drugs can be delivered lessfrequently provided plasma levels of the active moiety are sufficient tomaintain therapeutic effectiveness.

Plasma levels should reflect the potency of the drug. Generally, themore potent the compound the lower the plasma levels necessary toachieve efficacy.

V. Hydrophobic Pharmaceutical Agent Formulations

The formulations of the invention solubilize hydrophobic pharmaceuticalagents, such as quinazoline-, nitrothiazole-, and indolinone-basedcompounds. Because these pharmaceutical agents are typically insolublein aqueous environments, they require the addition of compounds that cansolubilize them before administration of the pharmaceutical agents to apatient.

For example, the preferred indolinone-based compound of the invention,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, has low solubilityin water (10 ng/mL), water miscible non-aqueous solvents, and oils. Anincrease in the indolinone-based compound's aqueous solubility inhydrotropic solute solutions like nicotinamide and pyridoxinehydrochloride was observed. It is soluble (15-20 mg/mL) in polarexcipients like polyethylene glycols of 300 and 400 MW. Among theseveral excipients studied, the indolinone-based compound hadcomparatively higher solubility (10-15 mg/mL) in aromatic solvents likebenzyl alcohol and in polyglycolized lipids (25-30 mg/mL) like LABRASOLand GELUCIRE.

The indolinone-based compound is a planar aromatic compound and isnon-ionizable in the pharmaceutically acceptable range. The pH of theaqueous media, therefore, did not influence its aqueous solubility andit could not be converted to any salt form. It is lipophilic, with atheoretical logP (octanol-water partition co-efficient) of 3.76 and alog of ratio of octanol to water solubility (individually determined) of5.6.

While not being limited by any particular mechanism of action, it isbelieved that the components of the formulations described herein bindto the hydrophobic regions of the pharmaceutical agents. Thisconsequently exposes the polar regions of the solubilizing components tothe solvent environment. This encapsulation of the pharmaceutical agentsrenders them soluble in aqueous environments.

The components of the formulations solubilize pharmaceutical agents inspecific concentrations depending on the concentration of thepharmaceutical agents in a formulation. Thus, the pharmaceutical agentsmay precipitate out of solution when the concentrations of theformulation components are outside the prescribed ranges set forthherein.

For both parenteral and oral formulations, the pharmaceutical agentswill likely precipitate out of solution when the concentration ofpolyoxyhydrocarbyl compound is not between 0.01 to 10 g/mL, and thesurfactant concentration is not between 0.01 to 10 g/mL. In someformulations, the pharmaceutical agents will likely precipitate out ofsolution when the ethanol concentration is not between 0.01 to 1 g/mLand/or when the benzyl alcohol concentration is not between 0.001 to 1g/mL. In oral formulations, the hydrophobic pharmaceutical agent willlikely precipitate out of solution when the polyglycolized lipidconcentration is not between 0.01 to 10 g/mL. In some oral formulations,the hydrophobic pharmaceutical agent will likely precipitate out ofsolution when the concentration of pharmaceutically acceptable oils isnot between 0.01 to 10 g/mL.

EXAMPLES

The examples below are not limiting and are merely representative ofvarious aspects and features of the present invention. The examplesdemonstrate methods of testing the solubility of the hydrophobicpharmaceutical agents in the formulations. In addition, the examplesillustrate preparation procedures for the formulations of the invention.

Example 1

Parenteral Formulations of Indolinone-based Compounds

The feasibility of developing different types of parenteral formulationsincluding cosolvent-surfactant based formulations, emulsion formulationsand liposome based formulations was studied. Based on the ease ofdevelopment and drug strength, a cosolvent based formulation was chosen.

Formulations were prepared for3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone. The compositionsfor three parenteral formulations that enhance the solubility of theindolinone-based compound are given in Table 3.

TABLE 3 Composition of Parenteral Formulations: 772-22, 772-69, 772-64Injection Concentrate, Concentration % w/v Excipients 772-22 772-69772-64 indolinone 5.0 6.0 5 (mg/mL) PEG-400 35 45 30 Cremophor EL 25 3040 Benzyl Alcohol 2 2 2 Ethanol 11.4 23 30 (anhydrous) Sterile Water addto 100 mL add to 100 mL add to 100 mL *all concentrations are in % w/v

Formulations 772-22, 772-69, and 772-64 were diluted with water in a 1:1ratio. In addition, the 772-22, 772-69, and 772-64 formulations werediluted in 0.45 WO saline, at 1:1, 1:2, and 1:3 ratios respectively,before final intravenous administration.

The 772-22, 772-69, and 772-64 formulations can be used for other waterinsoluble drugs, including other indolinone-based compounds and otherhydrophobic pharmaceutical agents of the invention, such as quinazoline-and nitrothiazole-based compounds. A drug even more hydrophobic thanthose disclosed herein would likely be solubilized by the formulation ofthe invention. The solubilization of a drug more hydrophobic than thosedisclosed herein could also be optimized by slightly modifying theamount of surfactant and/or polyethylene glycol concentrations.

In addition, the drug concentration can be increased or decreasedwithout modifying the contents of the formulations described herein. Theformulation composition can be slightly modified to accommodatesubstantial increases or decreases in drug concentration by, forexample, increasing or decreasing the concentration of surfactant and/orpolyethylene glycol. The concentrations of alcohols can also be modifiedin the formulation to accommodate different drug concentrations.

The preferred parenteral formulation is given in Table 4.

TABLE 4 Composition of IV formulation Excipients Concentration % w/vIndolinone 4.5 mg/mL PEG-400 45 CREMOPHOR EL ® 31.5 Benzyl alcohol 2Ethanol qs

Example 2

Parenteral Formulation Procedures

The protocol for the generic method used to prepare parenteralformulations of the invention is given below.

Generic Parenteral Formulation Preparation:

1. Weigh appropriate amounts of PEG-400.

2. Dissolve drug into PEG-400.

3. Add appropriate quantity of ethanol and benzyl alcohol to thePEG-400-drug solution. Mix by stirring and/or vortexing, depending onthe volume.

4. Weigh in an appropriate quantity surfactant.

5. Add distilled water to a total volume of 100 mL milliliters.

6. Mix and filter through a 0.2 μm nylon disposable filter unit(Nalgene) before use.

7. Store samples in the dark at temperatures equal to 25° C. or below.

The protocol for the method used to prepare a preferred parenteralformulation of the invention is given below.

772-22 Formulation Preparation:

1. Weigh 35 grams of PEG-400.

2. Dissolve appropriate amount of drug into PEG-400.

3. Add 10 grams of ethanol and 2 grams of benzyl alcohol to thePEG-400/drug solution. Mix by stirring followed by sonication for notmore than 2 minutes.

4. Weigh 25 grams of CREMOPHOR EL®.

5. Add distilled sterile water to a total volume of 100 mL milliliters.

6. Mix by stirring followed by sonication for more than two minutes.

7. Filter through a 0.2 Am disposable filter unit (Nalgene).

8. Store samples in the dark at temperatures equal to 25° C. or below.

Example 3

Oral Formulations of Indolinone-Based Compounds

Oral formulations were prepared for3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone. The compositionsfor three formulations that enhance the solubility of theindolinone-based compound are given (Table 5).

TABLE 5 Composition of Oral Formulations: 898-52, 698-99, 980-33Formulation 898-52 698-99 980-33 **Excipients Conc. % w/w Conc. % w/wConc. % w/w *Indolinone 3.0 3.0 3.0 GELUCIRE ® 44/14 70 0.0 0.0LABRASOL ® 0.0 76 70.0 Polyethylene 10 12 10 Glycol 600 Miglyol 812 10 010 CREMOPHOR, EL 10 12 10 *30 mg of the drug is added per gm of thevehicle **Either USP or NF grade excipients or high purity grade wereused

The 898-52, 698-99, 980-33 formulations can be used for other waterinsoluble drugs, including other indolinone-based compounds andhydrophobic pharmaceutical agents of the invention, includingquinazoline- and nitrothiazole-based compounds. For example, a drug evenmore hydrophobic than those disclosed herein would likely be solubilizedby the formulations of the invention. The solubilization of a drug morehydrophobic than those disclosed herein could also be optimized byslightly modifying the amount of surfactant, polyglycolized lipid, oil,and/or polyoxyhydrocarbyl concentrations.

In addition, the drug concentration can be increased or decreasedwithout modifying the contents of the formulations described herein. Theformulation composition can be slightly modified to accommodatesubstantial increases or decreases in drug concentration by, forexample, increasing or decreasing the concentration of surfactant,polyglycolized lipid, oil, and/or polyoxyhydrocarbyl compound.

Example 4

Oral Formulation Procedures

The protocol for the generic method used to prepare oral formulations ofthe invention is given below.

Generic Oral Preparation Procedure:

1. Melt GELUCIRE® 44/14 at 55-60° C. or heat LABRASOL® at 40° C.

2. Dissolve the other excipients into this melt.

3. Dissolve by stirring or sonicating, the hydrophobic pharmaceuticalagent into this molten mixture.

4. The liquid melt can be filled into either a hard gelatin capsule or asoft gelatin capsule or used as such. The GELUCIRE® based formulation isa semi-solid at room temperature. The LABRASOL® based formulation isliquid at room temperature.

5. To obtain a solid pharmaceutical composition, the liquefiedformulation (by heating if it is a semi-solid) is slowly mixed with agranulating agent until the mixture is a dry solid. This can be thenfilled into a hard gelatin capsule.

6. Store formulations in the dark at room temperature.

Example 5

Administration of Parenteral Formulations to Mammals Decreased TumorSize

Formulations of 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone havebeen administered to mice for pharmaco-kinetic studies. Formulations772-22, 772-69, and 772-64 were diluted with water or in 0.45% saline,at 1:1, and 1:2 ratios respectively, before final intravenousadministration.

Formulations of 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone in772-22 (50 μL, 100 μL) and 772-69 (50 μL) were also administered totumor-bearing mice without dilution by intraperitoneal injection. Themice were administered the formulation for more than 21 days.

The formulations were efficacious—the cancer tumor sizes decreased in alarge fraction of the mice tested in this study.

Example 6

Bioavailability Studies Including Administration of Oral Formulations toMammals

Several types of oral formulations, including formulations withmicronized drug substances with and without surfactants, soliddispersions, lipid solutions and self emulsifying lipid vehicles likeLABRASOL and GELUCIRE, have been evaluated. Among the many studied, thepolyglycolized lipid-based formulations showed higher bioavailabilities.Formulation approaches like micronization, solid dispersions andsolutions did not _enhance bioavailability.

Four groups of three beagle dogs were involved in the bioavailabilitystudy. All of the dogs were dosed intravenously at either 1 or 2 mg/kg.The absolute oral bioavailability of the oral formulations wasdetermined as compared to the intravenous dose at either 1 or 2 mg/kg.Groups 1 and 4 were dosed at 2 mg/kg at the beginning of the study andtheir PK parameters were determined again at the end of the study.Groups 2 and 3 were dosed at 1 mg/kg. All the candidate oralformulations were dosed at 50 mg/kg in hard gelatin capsules. Theformulations were melted at 60° C., filled into the capsule and storedat room temperature protected from light. The formulation is asemi-solid at room temperature. When dosed as a gavage, the formulationwas melted at 60° C. and cooled to 37.5° C. before dosing.

The formulations that were tested for bioavailability are given in FIG.1; the dosing regimens are given in FIG. 2.

One LABRASOL-based polyglycolized lipid formulation and one GELUCIRE44/14-based polyglycolized lipid formulation had bioavailabilities inbeagle dogs of 3±2% and 13±8, respectively. The LABRASOL formulationshowed efficacy in the preclinical mice model for angiogenesis. At 50mg/kg, the GELUCIRE-based formulation had about 45-fold higherbioavailability than the micronized formulation and about 4.5-foldhigher bioavailability as compared to the LABRASOL-based formulation.

Factors other than dissolution are affecting bioavailability as theLABRASOL formulation in which the drug was in solution both in theformulation and on dilution (1:100, 1:10) with water pH 6.5-7, had abioavailability lower than the semi-solid GELUCIRE-based formulation.

GELUCIRE-based formulation 898-52 provided the highest bioavailability,but there was variation between the dogs. Further testing of formulation898-52 at 15, 50 and 100 mg/kg in fasted dogs, 50 mg/kg in fed dogs, and50 mg/kg as a gavage (formulation melted and dosed at 37.5° C.) infasted dogs indicated that neither food nor form (liquid or solid) hadan effect on the bioavailability.

The excipients of formulation 898-52 were further tested for optimalbioavailability by: (a) substituting the polyglycolized lipid, GELUCIRE44/14, with the higher melting, and hence slower dissolving, GELUCIRE50/13; substituting GELUCIRE 44/14 with a mixture of LABRASOL andGELUCIRE 44/14; and substituting GELUCIRE 44/14 with a mixture ofGELUCIRE 44/14 and GELUCIRE 50/13; (b) substituting the oil, Miglyol812, with an MCM like Capmul; (c) substituting the surfactant, CREMOPHOREL with Polysorbate; (d) increasing the surfactant concentration; (e)adding lecithin, sodium lauryl sulfate, and sodium taurocholate; and (f)adding oleic acid. However, the optimization studies did not result inan increase in bio-availability over the original GELUCIRE formulation.

Analyses of Alanine aminotransferase and gamma glutamyl transpeptidasevalues were determined for all the dogs from base line up to cycle 13(every week). The Alanine aminotransferases and Gamma glutamyltranspeptidase values did not show any significant elevation over thetime span used for this study, nor any other treatment-related effect.

Example 6

Formulation Stability Studies Temperature Stability Studies

The temperature stability of formulation 898-52 (lot # 1035-049) at 25°C., 40° C. and 80° C. was tested. The two week stability profile of theformulation is shown in Table 6.

TABLE 6 Temperature Stability of the Indolinone Oral Formulation %Recovery Temp. Initial 6 days 2 weeks 25° C. 100% 100.49%  100.25%  40°C. 99.75% 99.54% 80° C. 98.44% 98.00%

pH-Stability

The pH stability profile of the indolinone-based parenteral formulation,772-69, was studied in the range of pH 2 to pH 9, at 5, 25, 40, 60 and80° C. The formulation at pH 9 (apparent) had the maximum stabilitybased on the degradation profile and indolinone-based compound recovery.The pH of this formulation on dilution with 0.45% saline or water rangedfrom 6.6-6.9. The pH-stability profile of the parenteral formulation,772-69, at 4 weeks is given in Table 7.

TABLE 7 pH-Stability of the Parenteral Formulation % INDOLINONERemaining at Temperatures (A/A) pH 5° C. 25° C. 40° C. 60° C. 80° C. 2100.7 100.0 87.6 88.9 78.1 3 100.6 100.0 88.4 90.3 79.9 4  97.7 100.089.4 88.6 78.2 5  96.3 100.0 93.4 92.4 82.3 6  99.3 100.0 97.3 94.6 88.57 102.4 100.0 100.5  96.2 92.4 9 104.4 100.0 102.9  97.9 92.5

One skilled in the art would readily appreciate that the presentinvention is well adapted to carry out the objects and obtain the endsand advantages mentioned, as well as those inherent therein. Themolecular complexes and the methods, procedures, treatments, molecules,specific compounds described herein are presently representative ofpreferred embodiments are exemplary and are not intended as limitationson the scope of the invention. Changes therein and other uses will occurto those skilled in the art which are encompassed within the spirit ofthe invention are defined by the scope of the claims.

It will be readily apparent to one skilled in the art that varyingsubstitutions and modifications may be made to the invention disclosedherein without departing from the scope and spirit of the invention.

All patents and publications mentioned in the specification areindicative of the levels of those skilled in the art to which theinvention pertains. All patents and publications are herein incorporatedby reference to the same extent as if each individual publication wasspecifically and individually indicated to be incorporated by reference.

The invention illustratively described herein suitably may be practicedin the absence of any element or elements, limitation or limitationswhich is not specifically disclosed herein. Thus, for example, in eachinstance herein any of the terms “comprising”, “consisting essentiallyof” and “consisting of” may be replaced with either of the other twoterms. The terms and expressions which have been employed are used asterms of description and not of limitation, and there is no intentionthat in the use of such terms and expressions of excluding anyequivalents of the features shown and described or portions thereof, butit is recognized that various modifications are possible within thescope of the invention claimed.

In particular, although the formulations described herein have beenidentified by the excipients added to the formulations, the invention ismeant to also cover the final formulation formed by the combination ofthese excipients. Specifically, the invention includes formulations inwhich one to all of the added excipients undergo a reaction duringformulation and are no longer present in the final formulation, or arepresent in modified forms.

In addition, where features or aspects of the invention are described interms of Markush groups, those skilled in the art will recognize thatthe invention is also thereby described in terms of any individualmember or subgroup of members of the Markush group. For example, if X isdescribed as selected from the group consisting of bromine, chlorine,and iodine, claims for X being bromine and claims for X being bromineand chlorine are fully described.

Other embodiments are within the following claims.

What is claimed is:
 1. A formulation comprising: (a) one or morehydrophobic pharmaceutical agents, wherein said agents are independentlyselected from the group consisting of quinazoline-, nitrothiazole-, andindolinone-based compounds; (b) one or more polyoxyhydrocarbylcompounds; and (c) one or more pharmaceutically acceptable surfactants;wherein said indolinone-based compound is an indolinone-based compoundof formula VI

at a concentration of about 0.1 to about 100 mg/ml; wherein R₁, R₂, R₃,R₄ are selected from the group consisting of hydrogen, trihalomethyl,hydroxyl, thioether, cyano, alkoxy, alkyl, amino, bromo, fluoro, chloro,iodo, mercapto, thio, cyanoamido, alkylthio, aryl, heteroaryl, carboxyl,ester, oxo, alkoxycarbonyl, alkenyl, alkoxy, nitro, alkoxyl, and amidomoieties; and R₅ is an optionally substituted aryl or heteroaryl cyclicmoiety; wherein at least one of said surfactants for theindolinone-based compound is PEG-400 at a concentration of about 0.01 toabout 10 g/ml and at least one of said polyoxyhydrocarbyl compounds forthe indolinone-based compound is ethoxylated castor oil at aconcentration of about 0.01 to 10 g/ml.
 2. The formulation of claim 1,wherein said hydrophobic pharmaceutical agent is a quinazoline-basedcompound of formula I,

wherein R₁, R₂, R₃, R₄, R₅ and R₆ are independently selected from thegroup consisting of: (i) hydrogen; (ii) saturated or unsaturated alkyl;(iii) an aryl optionally substituted with one, two, or threesubstituents independently selected from the group consisting of alkyl,alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester moieties;(iv) an amine of formula —NX₂X₃, wherein X₂ and X₃ are independentlyselected from the group consisting of hydrogen, saturated or unsaturatedalkyl, and homocyclic or heterocyclic ring moieties; (v) halogen ortrihalomethyl; (vi) a ketone of formula —CO—X₄, wherein X₄ is selectedfrom the group consisting of alkyl and homocyclic or heterocyclic ringmoieties; (vii) a carboxylic acid of formula —(X₅)_(n)—COOH or ester offormula —(X₆)_(n)—COO—X₇, wherein X₅, X₆, and X₇ and are independentlyselected from the group consisting of alkyl and homocyclic orheterocyclic ring moieties and wherein n is 0 or 1; (viii) an alcohol offormula (X₈)_(n)—OH or an alkoxy moiety of formula —(X₈)_(n)—O—X₉,wherein X₈ and X₉ are independently selected from the group consistingof saturated or unsaturated alkyl and homocyclic or heterocyclic ringmoieties, wherein said ring is optionally substituted with one or moresubstituents independently selected from the group consisting of alkyl,alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester andwherein n is 0 or 1; (ix) an amide of formula —NHCOX₁₀, wherein X₁₀ isselected from the group consisting of alkyl, hydroxyl, and homocyclic orheterocyclic ring moieties, wherein said ring is optionally substitutedwith one or more substituents independently selected from the groupconsisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, nitro,and ester; (x) —SO₂NX₁₁X₁₂, wherein X₁₁ and X₁₂ are selected from thegroup consisting of hydrogen, alkyl, and homocyclic or heterocyclic ringmoieties; (xi) a homocyclic or heterocyclic ring moiety optionallysubstituted with one, two, or three substituents independently selectedfrom the group consisting of alkyl, alkoxy, halogen, trihalomethyl,carboxylate, nitro, and ester moieties; (xii) an aldehyde of formula—CO—H; (xiii) a sulfone of formula —SO₂—X₁₃, wherein X₁₃ is selectedfrom the group consisting of saturated or unsaturated alkyl andhomocyclic or heterocyclic ring moieties; and (xiv) a nitro of formula—NO₂.
 3. The formulation of claim 2, wherein said quinazoline-basedcompound is 4-(3-Bromophenyl)-6,7-dimethoxyquinazoline.
 4. Theformulation of claim 1, wherein said hydrophobic pharmaceutical agent isa nitrothiazole-based compound of formula IV,

wherein R₁, R₂, and R₃ are independently selected from the groupconsisting of (i) hydrogen; (ii) saturated or unsaturated alkyl; (iii)an aryl optionally substituted with one, two, or three substituentsindependently selected from the group consisting of alkyl, alkoxy,halogen, trihalomethyl, carboxylate, nitro, and ester moieties; (iv) anamine of formula —NX₂X₃, wherein X₂ and X₃ are independently selectedfrom the group consisting of hydrogen, saturated or unsaturated alkyl,and homocyclic or heterocyclic ring moieties; (v) halogen ortrihalomethyl; (vi) a ketone of formula —CO—X₄, wherein X₄ is selectedfrom the group consisting of alkyl and homocyclic or heterocyclic ringmoieties; (vii) a carboxylic acid of formula —(X₅)_(n)—COOH or ester offormula —(X₆)_(n)—COO—X₇, wherein X₅, X₆, and X₇ and are independentlyselected from the group consisting of alkyl and homocyclic orheterocyclic ring moieties and wherein n is 0 or 1; (viii) an alcohol offormula (X₈)_(n)—OH or an alkoxy moiety of formula —(X₈)_(n)—O—X₉,wherein X₈ and X₉ are independently selected from the group consistingof saturated or unsaturated alkyl and homocyclic or heterocyclic ringmoieties, wherein said ring is optionally substituted with one or moresubstituents independently selected from the group consisting of alkyl,alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester andwherein n is 0 or (ix) an amide of formula —NHCOX₁₀, wherein X₁₀ isselected from the group consisting of alkyl, hydroxyl, and homocyclic orheterocyclic ring moieties, wherein said ring is optionally substitutedwith one or more substituents independently selected from the groupconsisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, nitro,and ester; (x) —SO₂NX₁₁X₁₂, wherein X₁₁ and X₁₂ are selected from thegroup consisting of hydrogen, alkyl, and homocyclic or heterocyclic ringmoieties; (xi) a homocyclic or heterocyclic ring moiety optionallysubstituted with one, two, or three substituents independently selectedfrom the group consisting of alkyl, alkoxy, halogen, trihalomethyl,carboxylate, nitro, and ester moieties; (xii) an aldehyde of formula—CO—H; (xiii) a sulfone of formula —SO₂—X₁₃, wherein X₁₃ is selectedfrom the group consisting of saturated or unsaturated alkyl andhomocyclic or heterocyclic ring moieties; and (xiv) a nitro of formula—NO₂.
 5. The formulation of claim 1, wherein said hydrophobicpharmaceutical agent is a nitrothiazole-based compound of formula V,

wherein R₁ and R₂ are independently selected from the group consistingof: (i) hydrogen; (ii) saturated or unsaturated alkyl; (iii) an aryloptionally substituted with one, two, or three substituentsindependently selected from the group consisting of alkyl, alkoxy,halogen, trihalomethyl, carboxylate, nitro, and ester moieties; (iv) anamine of formula —NX₂X₃, wherein X₂ and X₃ are independently selectedfrom the group consisting of hydrogen, saturated or unsaturated alkyl,and homocyclic or heterocyclic ring moieties; (v) halogen ortrihalomethyl; (vi) a ketone of formula —CO—X₄, wherein X₄ is selectedfrom the group consisting of alkyl and homocyclic or heterocyclic ringmoieties; (vii) a carboxylic acid of formula —(X₅)_(n)—COOH or ester offormula —(X₆)_(n)—COO—X₇, wherein X₅, X₆, and X₇ and are independentlyselected from the group consisting of alkyl and homocyclic orheterocyclic ring moieties and wherein n is 0 or 1; (viii) an alcohol offormula (X₈)_(n)—OH or an alkoxy moiety of formula —(X₈)_(n)—O—X₉,wherein X₈ and X₉ are independently selected from the group consistingof saturated or unsaturated alkyl and homocyclic or heterocyclic ringmoieties, wherein said ring is optionally substituted with one or moresubstituents independently selected from the group consisting of alkyl,alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester andwherein n is 0 or 1; (ix) an amide of formula —NHCOX₁₀, wherein X₁₀ isselected from the group consisting of alkyl, hydroxyl, and homocyclic orheterocyclic ring moieties, wherein said ring is optionally substitutedwith one or more substituents independently selected from the groupconsisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, nitro,and ester; (x) —SO₂NX₁₁X₁₂, wherein X₁₁ and X₁₂ are selected from thegroup consisting of hydrogen, alkyl, and homocyclic or heterocyclic ringmoieties; (xi) a homocyclic or heterocyclic ring moiety optionallysubstituted with one, two, or three substituents independently selectedfrom the group consisting of alkyl, alkoxy, halogen, trihalomethyl,carboxylate, nitro, and ester moieties; (xii) an aldehyde of formula—CO—H; (xiii) a sulfone of formula —SO₂—X₁₃, wherein X₁₃ is selectedfrom the group consisting of saturated or unsaturated alkyl andhomocyclic or heterocyclic ring moieties; and (xiv) a nitro of formula—NO₂.
 6. The formulation of claim 1, wherein said one or morepolyoxyhydrocarbyl compounds are water soluble polymers selected fromthe group consisting of: water soluble carbohydrates, water solublecarbohydrate derivatives, water soluble mixed oxyalkylene polymers, andthe polymeric form of ethylene glycol.
 7. The formulation of claim 6,wherein said one or more polyoxyhydrocarbyl compounds are poly(ethyleneglycol) (PEG) or PEG derivatives.
 8. The formulation of claim 1, whereinsaid one or more surfactants are one or more non-ionic surfactants. 9.The formulation of claim 1, wherein said one or more surfactants areindependently selected from the group consisting of: polyoxyethylenesorbitan fatty acid esters, glyceryl monooleate, polyvinyl alcohol,ethylene oxide copolymers, polyol moieties, and sorbitan esters.
 10. Theformulation of claim 1, wherein said one or more surfactants are one ormore ethoxylated castor oils.
 11. The formulation of claim 1, whereinsaid formulation further comprises one or more pharmaceuticallyacceptable alcohols.
 12. The formulation of claim 11, wherein said oneor more alcohols are independently selected from the group consisting ofethanol, benzyl alcohol, propylene glycol, 2-(2-ethoxyethoxy)ethanol,and glycerol.
 13. The formulation of claim 12, wherein said alcohols areindependently selected from ethanol and benzyl alcohol.
 14. Theformulation of any of claims 1, 6-10, 11-13, wherein said formulation iseffective in treating or preventing a cell proliferative disorder in apatient in need of such treatment.
 15. The formulation of claim 14,wherein said formulation is administered parenterally.
 16. Theformulation of claim 15, wherein said patient is a mammal and said cellproliferative disorder is selected from the group consisting of cancers,blood vessel proliferative disorders, fibrotic disorders, and autoimmunedisorders.
 17. A method of preventing or treating a cell proliferativedisorder in a patient in need of treatment comprising the followingsteps: (a) diluting a parenteral formulation into a pharmaceuticallyacceptable solution, said parenteral formulation comprising one or morehydrophobic pharmaceutical agents, wherein said agents are independentlyselected from the group consisting of a quinazolin-, a nitrothiazole-and an indolinone-based compound; one or more polyoxyhydrocarbylcompounds; and one or more pharmaceutically acceptable surfactants;wherein said indolinone-based compound is an indolinone-based compoundof formula VI

at a concentration of about 0.1 to about 100 mg/ml; wherein R₁, R₂, R₃,R₄ are selected from the group consisting of hydrogen, trihalomethyl,hydroxyl, thioether, cyano, alkoxy, alkyl, amino, bromo, fluoro, chloro,iodo, mercapto, thio, cyanoamido, alkylthio, aryl, heteroaryl, carboxyl,ester, oxo, alkoxycarbonyl, alkenyl, alkoxy, nitro, alkoxyl, and amidomoieties; and R₅ is an optionally substituted aryl or heteroaryl cyclicmoiety; wherein at least one or said surfactants for theindolinone-based compound is PEG-400 at a concentration of about 0.01 toabout 10 g/ml and at least one of said polyoxyhydrocarbyl compounds forthe indolinone-based compound is ethoxylated castor oil at aconcentration of about 0.01 to 10 g/ml; and (b) parenterallyadministering the diluted formulation to the patient.
 18. The method ofclaim 17, wherein said formulation further comprises one or morepharmaceutically acceptable alcohols.
 19. The method of either claim 17or 18, wherein said patient is a mammal and said is cell proliferativedisorder selected from the group consisting of cancers, blood vesselproliferative disorders, fibrotic disorders, and autoimmune disorders.20. A formulation comprising: (a) one or more hydrophobic pharmaceuticalagents, wherein said agents are independently selected from the groupconsisting of quinazoline-, nitrothiazole-, and indolinone-basedcompounds; (b) one or more polyoxyhydrocarbyl compounds; (c) one or morepharmaceutically acceptable surfactants; wherein said indolinone-basedcompound is an indolinone-based compound of formula VI

at a concentration of about 5 mg/ml, wherein R₁, R₂, R₃, R₄ are selectedfrom the group consisting of hydrogen, trihalomethyl, hydroxyl,thioether, cyano, alkoxy, alkyl, amino, bromo, fluoro, chloro, iodo,mercapto, thio, cyanoamido, alkylthio, aryl, heteroaryl, carboxyl,ester, oxo, alkoxycarbonyl, alkenyl, alkoxy, nitro, alkoxyl, and amidomoieties; and R₅ is an optionally substituted aryl or heteroaryl cyclicmoiety; wherein at least one of said surfactant for the indolinone-basedcompounds is PEG-400 at a concentration of about 0.35 g/ml and at leastone of said polyoxyhydrocarbyl compounds for the indolinone-basedcompound is ethoxylated castor oil at a concentration of about 0.25g/ml.
 21. The formulation of claims 20, wherein said indolinone-basedcompound is selected from the group consisting of3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-amino)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-chloro)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, and3-[(3-methylthiophenyl-5-yl)methylene]-(4-methyl)-2-indolinone.
 22. Theformulation of claim 21, wherein said indolinone-based compound is3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone.
 23. A formulationcomprising: (a) an indolinone-based compound of formula VI

at a concentration of about 6 mg/ml; (b) one or more polyoxyhydrocarbylcompounds; and (c) one or more pharmaceutically acceptable surfactants;wherein R₁, R₂, R₃, R₄ are selected from the group consisting ofhydrogen, trihalomethyl, hydroxyl, thioether, cyano, alkoxy, alkyl,amino, bromo, fluoro, chloro, iodo, mercapto, thio, cyanoamido,alkylthio, aryl, heteroaryl, carboxyl, ester, oxo, alkoxycarbonyl,alkenyl, alkoxy, nitro, alkoxyl, and amido moieties; and R₅ is anoptionally substituted aryl or heteroaryl cyclic moiety; and R₅ is anoptionally substituted aryl or heteroaryl cyclic moiety; wherein atleast one of said surfactant is PEG-400 at a concentration of about 0.45g/ml and at least one of said polyoxyhydrocarbyl compounds isethoxylated castor oil at a concentration of about 0.30 g/ml.
 24. Theformulation of claim 22, wherein said indolinone-based compound isselected from the group consisting of3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-amino)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-(5-chloro)-2-indolinone,3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, and3-[(3-methylthiophenyl-5-yl)methylene]-(4-methyl)-2-indolinone.
 25. Theformulation of claim 24, wherein said indolinone-based compound is3-[2,4-dimenthylpyrrol-5-yl)methylene]-2-indolinone.